Method of treating second and third degree burns using oxidative reductive potential water solution

ABSTRACT

A method of treating burns, preferably second and third degree burns, by administration of an oxidative reduction potential (ORP) water solution that is stable for at least twenty-four hours is provided.

CROSS-REFERENCE TO RELATED APPLICATIONS

This patent application claims the benefit of U.S. Provisional PatentApplication Nos. 60/760,635 filed Jan. 20, 2006; 60/760,567 filed Jan.20, 2006; 60/760,645 filed Jan. 20, 2006; 60/760,557 filed Jan. 20,2006; 60/730,743 filed Oct. 27, 2005; 60/676,883 filed May 2, 2005;60/667,101 filed Mar. 31, 2005; and 60/664,361, filed Mar. 23, 2005;each of which is hereby incorporated herein in their entirety byreference.

FIELD OF THE INVENTION

This invention pertains to a method of treating burns, preferably secondand third degree burns, by administration of oxidative reductivepotential water solutions.

BACKGROUND OF THE INVENTION

Oxidative reductive potential (ORP) water, also known as super-oxidizedwater, can be used as a non-toxic disinfectant to eradicatemicroorganisms, including bacteria, viruses and spores, in variety ofsettings. For example, ORP water may be applied in the healthcare andmedical device fields to disinfect surfaces and medical equipment.Advantageously, ORP water is environmentally safe and, thus, avoids theneed for costly disposal procedures. ORP water also has application inwound care, medical device sterilization, food sterilization, hospitals,consumer households and anti-bioterrorism.

Although ORP water is an effective disinfectant, it has an extremelylimited shelf-life, usually only a few hours. As a result of this shortlifespan, the production of ORP water must take place in close proximityto where ORP water is to be used as a disinfectant. This means that ahealthcare facility, such as a hospital, must purchase, house andmaintain the equipment necessary to produce ORP water. Additionally,prior manufacturing techniques have not been able to produce sufficientcommercial-scale quantities of ORP water to permit its widespread use asa disinfectant at healthcare facilities.

Accordingly, a need exists for an ORP water that is stable over anextended period of time and methods of using such an ORP water. A needalso exists for cost-effective methods of preparing commercial-scalequantities of ORP water. The present invention provides such an ORPwater and methods of preparing and using such an ORP water.

ORP water has also been used as a tissue cell growth promoter inpatients as described in U.S. Patent Application Publication2002/0160053 A1. Infections remain a problem in wound care especiallywith the emergence of multi-antibiotic resistant bacteria. Suchinfections include, for example, Acinetobacter baumannii, Staph aureus,Ps. aeruginosa, E. coli, and others. Accordingly, a need exists forcompositions containing ORP water for use in the treatment of burns thatprevent infections. These and other advantages of the invention, as wellas additional inventive features, will be apparent from the descriptionof the invention provided herein.

BRIEF SUMMARY OF THE INVENTION

The present invention provides a method of treating burns in a patientby administering an oxidative reductive potential (ORP) water solution,wherein the solution is stable for at least twenty-four hours. Theinvention also is directed to a method of treating burns in a patient byadministering an oxidative reductive potential water solution, whereinthe solution comprises anode water and cathode water. In one embodiment,the ORP water solution used in the method of the invention comprises oneor more chlorine species.

The present invention additionally provides a method of treatingimpaired or damaged tissue, which method comprises contacting theimpaired or damaged tissue with a therapeutically effective amount of anORP water solution, wherein the solution is stable for at leasttwenty-four hours. The method includes treating tissue, which has beenimpaired or damaged by surgery or which has been impaired or damaged bycauses that are not necessarily relate to surgery, e.g., burns, cuts,abrasions, scrapes, rashes, ulcers, puncture wounds, infections, and thelike.

The present invention further provides a method of disinfecting asurface, which method comprises contacting the surface with ananti-infective amount of an ORP water solution, wherein the solution isstable for at least twenty-four hours. The surface can be biological,inanimate, or a combination of such surfaces can be disinfected inaccordance with the present invention. Biological surfaces include, e.g,muscle tissue, bone tissue, organ tissue, mucosal tissue, andcombinations thereof, can be disinfected in accordance with the presentinvention. Inanimate surfaces include, e.g., surgically implantabledevices, prosthetic devices, and medical devices.

Another aspect of the present invention includes a formulation fortopical administration comprising an oxidative reductive potential watersolution and a thickening agent, wherein the formulation is stable forat least twenty-four hours.

The invention also pertains to a pharmaceutical dosage form comprising(1) a formulation for topical administration comprising an oxidativereductive potential water solution and a thickening agent and (2) asealed container, wherein the formulation is stable for at leasttwenty-four hours.

Additionally, the invention is directed to a method for treating acondition in a patient comprising topically administering to a patient atherapeutically effective amount of a formulation comprising anoxidative reductive potential solution and a thickening agent, whereinthe formulation is stable for at least about twenty-four hours.

The invention further provides a method for promoting wound healing in apatient comprising applying to a wound a formulation comprising anoxidative reductive potential water solution and a thickening agent,wherein the formulation is administered in an amount sufficient topromote wound healing, and wherein the formulation is stable for atleast about twenty-four hours.

The invention additionally provides a method for preventing a conditionin a patient comprising topically administering to a patient atherapeutically effective amount of a formulation comprising anoxidative reductive potential water solution and a thickening agent,wherein the formulation is stable for at least about twenty-four hours.

Another aspect of the present invention includes an apparatus forproducing an oxidative reductive potential water solution comprising atleast two electrolysis cells, wherein each cell comprises an anodechamber, cathode chamber and salt solution chamber located between theanode and cathode chambers, wherein the anode chamber is separated fromthe salt solution chamber by an anode electrode and a first membrane,and the cathode chamber is separated from the salt solution chamber by acathode electrode and a second membrane. The apparatus may include arecirculation system for the salt solution supplied to the salt solutionchamber to permit the concentration of salt ions to be controlled andmaintained.

The invention further provides a process for producing oxidativereductive potential water solution comprising providing at least twoelectrolysis cells, wherein each cell comprises an anode chamber,cathode chamber and salt solution chamber located between the anode andcathode chambers, wherein the anode chamber is separated from the saltsolution chamber by an anode electrode and a first membrane, and thecathode chamber is separated from the salt solution chamber by a cathodeelectrode and a second membrane, providing a flow of water through theanode chamber and cathode chamber, providing a flow of a salt solutionthrough the salt solution chamber, providing electrical current to theanode electrode and cathode electrode simultaneously with the flow ofwater through the anode and cathode chambers and the flow of saltsolution through the salt solution chamber, and collecting the oxidativereductive potential water solution produced by the electrolysis cells.

The invention is also directed to a process for producing oxidativereductive potential water solution comprising providing at least oneelectrolysis cell, wherein the cell comprises an anode chamber, cathodechamber and salt solution chamber located between the anode and cathodechambers, wherein the anode chamber is separated from the salt solutionchamber by an anode electrode and a first membrane, and the cathodechamber is separated from the salt solution chamber by a cathodeelectrode and a second membrane, providing a flow of water through theanode chamber and cathode chamber, providing a flow of salt solutionthrough the salt solution chamber, providing electrical current to theanode electrode and cathode electrode simultaneously with the flow ofwater through the anode and cathode chambers and the flow of saltsolution through the salt solution chamber, and collecting the oxidativereductive potential water produced by the electrolysis cell, wherein thesolution comprises anode water and cathode water.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic diagram of a three-chambered electrolysis cell forproducing an oxidative reductive potential water solution of the presentinvention.

FIG. 2 illustrates a three-chambered electrolysis cell and depicts ionicspecies generated therein.

FIG. 3 is a schematic flow diagram of a process for producing anoxidative reductive potential water of the present invention.

FIGS. 4A-4C depicts a graphical comparison of cell viability, apoptosisand necrosis in human dermal fibroblasts (HDFs) treated with anexemplary ORP water solution (MCN) versus hydrogen peroxide (HP).

FIG. 5 is a graphical comparison of the levels of8-hydroxy-2′-deoxiguanosine (8-OHdG) adducts in HDFs treated with anexemplary ORP water solution (MCN) versus 500 μM hydrogen peroxide (HP).

FIGS. 6A-6B illustrate the expression of a senescence associated withβ-galactosidase in HDFs after chronic exposure to low concentrations ofan exemplary ORP water solution (MCN) versus hydrogen peroxide (HP).

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a method of preventing or treating acondition in a patient, which method comprises administering to thepatient a therapeutically effective amount of an oxidative reductivepotential (ORP) water solution, wherein the solution is stable for atleast twenty-four hours. The condition can include, e.g., medicalconditions, illnesses, injuries, allergies, and the like, which aretreatable with the ORP water solution of the present invention.

The therapeutically effective amount administered to the patient, e.g.,an animal, particularly a human, in the context of the present inventionshould be sufficient to effect a therapeutic or prophylactic response inthe patient over a reasonable time frame. The dose can be readilydetermined using methods that are well known in the art. One skilled inthe art will recognize that the specific dosage level for any particularpatient will depend upon a variety of factors. For example, the dose canbe determined based on the strength of the particular ORP water solutionemployed, the severity of the condition, the body weight of the patient,the age of the patient, the physical and mental condition of thepatient, general health, sex, diet, and the like. The size of the dosealso can be determined based on the existence, nature, and extent of anyadverse side effects that might accompany the administration of aparticular ORP water solution. It is desirable, whenever possible, tokeep adverse side effects to a minimum.

Factors, which can be taken into account for a specific dosage caninclude, for example, bioavailability, metabolic profile, time ofadministration, route of administration, rate of excretion,pharmacodynamics associated with a particular ORP water solution in aparticular patient, and the like. Other factors can include, e.g., thepotency or effectiveness of the ORP water solution with respect to theparticular condition to be treated, the severity of the symptomspresented prior to or during the course of therapy, and the like. Insome instances, what constitutes a therapeutically effective amount alsocan be determined, in part, by the use of one or more of the assays,e.g., bioassays, which are reasonably clinically predictive of theefficacy of a particular ORP water solution for the treatment orprevention of a particular condition.

The ORP water solution of the present invention can be administeredtherapeutically, alone or in combination with one or more othertherapeutic agents, to a patient, e.g., a human, e.g., to treat anexisting condition. The ORP water solution of the present invention alsocan be administered prophylactically, alone or in combination with oneor more other therapeutic agents, to a patient, e.g., a human, that hasbeen exposed to one or more causative agents associated with thecondition. For example, the ORP water solution of the invention can besuitably administered to a patient that has been exposed to one or moreinfection-causing microorganisms (e.g., viruses, bacteria and/or fungi)prophylactically to inhibit or decrease the likelihood of infection in apatient, or decrease the severity of an infection that develops as aresult of such exposure.

One skilled in the art will appreciate that suitable methods ofadministering the ORP water solution of the present invention areavailable, and, although more than one route of administration can beused, a particular route can provide a more immediate and more effectivereaction than another route. The therapeutically effective amount can bethe dose necessary to achieve an “effective level” of the ORP watersolution in an individual patient. The therapeutically effective amountcan be defined, for example, as the amount required to be administeredto an individual patient to achieve a blood level, tissue level, and/orintracellular level of the ORP water of the present invention to preventor treat the condition in the patient.

When the effective level is used as a preferred endpoint for dosing, theactual dose and schedule can vary depending, for example, uponinterindividual differences in pharmacokinetics, distribution,metabolism, and the like. The effective level also can vary when the ORPwater solution of the present invention is used in combination with oneor more therapeutic agents other than the ORP water solution of thepresent invention, e.g., one or more anti-infective agents, one or more“moderating,” “modulating” or “neutralizing agents,” e.g., as describedin U.S. Pat. Nos. 5,334,383 and 5,622,848, one or more anti-inflammatoryagents, and the like.

An appropriate indicator can be used for determining and/or monitoringthe effective level. For example, the effective level can be determinedby direct analysis (e.g., analytical chemistry) or by indirect analysis(e.g., with clinical chemistry indicators) of appropriate patientsamples (e.g., blood and/or tissues). The effective level also can bedetermined, for example, by direct or indirect observations such as,e.g., the concentration of urinary metabolites, changes in markersassociated with the condition (e.g., viral count in the case of a viralinfection), decrease in the symptoms associated with the conditions, andthe like.

The ORP water of the present invention can be administered using anysuitable method of administration known in the art. The ORP water of thepresent invention can be administered in combination with one or morepharmaceutically acceptable carriers, vehicles, adjuvants, excipients,or diluents, which are known in the art. One skilled in the art caneasily determine the appropriate formulation and method ofadministration for administering the ORP water in accordance with thepresent invention. Any necessary adjustments in dose can be readily madeby a skilled practitioner to address the nature or severity of thecondition being treated in view of other factors, such as, e.g., sideeffects, changes in the patient's overall condition, and the like.

The ORP solution of the present invention can be administered to theupper airway as a steam or a spray. In addition, the ORP water solutionof the present invention can be administered by aerosolization,nebulization or atomization. When the ORP water solution of theinvention is administered by aerosolization, nebulization oratomization, it is preferably administered in the form of dropletshaving a diameter in the range of from about 1 micron to about 10microns.

Methods and devices, which are useful for aerosolization, nebulizationand atomization, are well known in the art. Medical nebulizers, forexample, have been used to deliver a metered dose of a physiologicallyactive liquid into an inspiration gas stream for inhalation by arecipient. See, e.g., U.S. Pat. No. 6,598,602. Medical nebulizers canoperate to generate liquid droplets, which form an aerosol with theinspiration gas. In other circumstances medical nebulizers may be usedto inject water droplets into an inspiration gas stream to provide gaswith a suitable moisture content to a recipient, which is particularlyuseful where the inspiration gas stream is provided by a mechanicalbreathing aid such as a respirator, ventilator or anaesthetic deliverysystem.

An exemplary nebulizer is described, for example, in WO 95/01137, whichdescribes a hand held device that operates to eject droplets of amedical liquid into a passing air stream (inspiration gas stream), whichis generated by a recipient's inhalation through a mouthpiece. Anotherexample can be found in U.S. Pat. No. 5,388,571, which describes apositive-pressure ventilator system which provides control andaugmentation of breathing for a patient with respiratory insufficiencyand which includes a nebulizer for delivering particles of liquidmedication into the airways and alveoli of the lungs of a patient. U.S.Pat. No. 5,312,281 describes an ultrasonic wave nebulizer, whichatomizes water or liquid at low temperature and reportedly can adjustthe size of mist. In addition, U.S. Pat. No. 5,287,847 describes apneumatic nebulizing apparatus with scalable flow rates and outputvolumes for delivering a medicinal aerosol to neonates, children andadults. Further, U.S. Pat. No. 5,063,922 describes an ultrasonicatomizer.

The method of the present invention also can be used for the preventionor treatment of an infection, which is treatable with the ORP watersolution of the present invention. The infection can be caused by one ormore infectious pathogens such as, for example, infectiousmicroorganisms. Such microorganisms can include, for example, viruses,bacteria, and fungi. The viruses can include, e.g., one or more virusesselected from the group consisting of adenoviruses, HIV, rhinoviruses,and flu viruses. The bacteria can include, e.g., one or more bacteriaselected from the group consisting of Escherichia coli, Pseudomonasaeruginosa, Staphylococcus aureus, and Mycobaterium tuberculosis. Thefungi can include, e.g., one or more fungi selected from the groupconsisting of Candida albicans, Bacillus subtilis and Bacillusathrophaeus. The method of the present invention also can be used forthe prevention or treatment of inflammatory conditions or allergicreactions, which are treatable with the ORP water solution of theinvention.

In addition, organisms that can be controlled, reduced, killed oreradicated by treatment with the ORP water solution used in accordancewith the invention include, e.g., Pseudomonas aeruginosa, Escherichiacoli, Enterococcus hirae, Acinetobacter baumannii, Acinetobacterspecies, Bacteroides fragilis, Enterobacter aerogenes, Enterococcusfaecalis, Vancomycin resistant-Enterococcus faecium (VRE, MDR),Haemophilus influenzae, Klebsiella oxytoca, Klebsiella pneumoniae,Micrococcus luteus, Proteus mirabilis, Serratia marcescens,Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcushaemolyticus, Staphylococcus hominis, Staphylococcus saprophyticus,Streptococcus pneumoniae, Streptococcus pyogenes, Salmonellacholeraesuis, Shigella dysenteriae, and other susceptible bacteria, aswell as yeasts, e.g., Trichophyton mentagrophytes, Candida albicans andCandida tropicalis. The ORP water solution can also be used inaccordance with the invention to control, reduce, kill or eradicateviruses including, e.g., adenovirus, human immunodeficiency virus (HIV),rhinovirus, influenza (e.g., influenza A), hepatitis (e.g., hepatitisA), coronavirus (responsible for, e.g., Severe Acute RespiratorySyndrome (SARS)), rotavirus, avian flu virus, respiratory syncytialvirus, herpes simplex virus, varicella zoster virus, rubella virus, andother susceptible viruses.

In another embodiment, the method of the present invention comprisesparenterally administering the ORP water solution of the invention.Parenteral administration can include administering the ORP watersolution of the invention intravenously, subcutaneously,intramuscularly, or intraperitoneally. In a preferred embodiment, theORP water solution of the present invention is administeredintravenously to prevent or treat a condition in accordance with themethod of the present invention. Suitable conditions can include, e.g.,viral myocarditis, multiple sclerosis, and AIDS. See, e.g., U.S. Pat.Nos. 5,334,383 and 5,622,848, which describe methods of treating viralmyocarditis, multiple sclerosis, and AIDS via intravenous administrationof ORP water solutions.

The present invention additionally provides a method of treatingimpaired or damaged tissue, which method comprises contacting theimpaired or damaged tissue with a therapeutically effective amount ofthe ORP water solution of the present invention. Any suitable method canbe used for contacting the impaired or damaged tissue, so as to treatthe impaired or damaged tissue in accordance with the present invention.For example, the impaired or damaged tissue can be treated in accordancewith the invention by irrigating the tissue with the ORP water solutionof the invention, so as to contact the impaired or damaged tissue withthe ORP water. Alternatively (and additionally), the ORP water solutionof the present invention can be administered as a steam or a spray, orby aerosolization, nebulization or atomization, as described herein, soas to contact the impaired or damaged tissue with the ORP water.

The method of the present invention can be used in the treatment oftissues, which have been impaired or damaged, e.g., by surgery. Forinstance, the method of the present invention can be used for treatingtissues, which have been impaired or damaged by an incision. Inaddition, the method of the present invention can be used for treatingtissues, which have been impaired or damaged by oral surgery, graftsurgery, implant surgery, transplant surgery, cauterization, amputation,radiation, chemotherapy, and combinations thereof. The oral surgery caninclude, for example, dental surgery such as, e.g., root canal surgery,tooth extraction, gum surgery, and the like.

The method of the present invention also includes treating tissues,which have been impaired or damaged by one or more burns, cuts,abrasions, scrapes, rashes, ulcers, puncture wounds, combinationsthereof, and the like, which are not necessarily caused by surgery. Themethod of the present invention also can be used for treating impairedor damaged tissue, which is infected, or tissue impaired or damaged dueto infection. Such infection can be caused by one or more infectiouspathogens, such as, e.g., one or more microorganisms selected from thegroup consisting of viruses, bacteria, and fungi, as described herein.

The present invention further provides a method of disinfecting asurface, which method comprises contacting the surface with ananti-infective amount of the ORP water solution of the presentinvention. In accordance with the method of the present invention, thesurface can be contacted using any suitable method. For example, thesurface can be contacted by irrigating the surface with the ORP watersolution of the invention, so as to disinfect the surface in accordancewith the invention. Additionally, the surface can be contacted byapplying the ORP water solution of the present invention to the surfaceas a steam or a spray, or by aerosolization, nebulization oratomization, as described herein, so as to disinfect the surface inaccordance with the invention. Further, the ORP water solution of thepresent invention can be applied to the surface with a cleaning wipe, asdescribed herein. By disinfecting a surface in accordance with thepresent invention, the surface may be cleansed of infectiousmicroorganisms. Alternatively (or additionally), the ORP water solutionof the present invention can be applied to the surface to provide abarrier to infection, thereby disinfecting a surface in accordance withthe present invention.

The method of the present invention can be used for disinfecting asurface, which is biological, inanimate, or a combination thereof.Biological surfaces can include, for example, tissues within one or morebody cavities such as, for example, the oral cavity, the sinus cavity,the cranial cavity, the abdominal cavity, and the thoracic cavity.Tissues within the oral cavity include, e.g., mouth tissue, gum tissue,tongue tissue, and throat tissue. The biological tissue also can includemuscle tissue, bone tissue, organ tissue, mucosal tissue, andcombinations thereof. Inanimate surfaces include, for example,surgically implantable devices, prosthetic devices, and medical devices.In accordance with the method of the present invention, the surfaces ofinternal organs, viscera, muscle, and the like, which may be exposedduring surgery, can be disinfected, e.g., to maintain sterility of thesurgical environment.

The present invention also provides formulations for topicaladministration comprising an oxidative reductive potential (ORP) watersolution and a thickening agent which are prepared to provide enhancedefficacy and stability.

The amount of water present the formulations of the invention isgenerally from about 10% by weight to about 95% by weight, based on theweight of the formulation. Preferably, the amount of water present isfrom about 50% by weight to about 90% by weight.

The formulations of the invention preferably include an ORP watersolution comprising anode water and cathode water. Anode water isproduced in the anode chamber of the electrolysis cell used in thepresent invention. Cathode water is produced in the cathode chamber ofthe electrolysis cell.

The formulation for topical administration according to the presentinvention further comprises a thickening agent. Any suitable thickeningagent may be used to produce a formulation having the desired viscositywhich is generally greater than the ORP water solution alone. Thethickening agent utilized is compatible with the ORP water solution andother optional components in the formulation. Suitable thickening agentsinclude, but are not limited to, polymers and hydroxyethylcellulose.Suitable polymers may be homopolymers or copolymers and are optionallycrosslinked. Other suitable thickening agents are generally known in art(see, e.g., Handbook of Cosmetic and Personal Care Additives, 2nd ed.,Ashe et al. eds. (2002), and Handbook of Pharmaceutical Excipients, 4thed., Rowe et al. eds. (2003)).

Preferred thickening agents are acrylic acid-based polymers. Morepreferably, the thickening agents are high molecular weight,crosslinked, acrylic acid-based polymers. These polymers have thefollowing general structure:

Such polymers are sold under the tradename Carbopol® by Noveon.Carbopol® polymers are generally supplied as rheology modifiers for usethickeners, suspending agents, and stabilizers in a variety of personalcare products, pharmaceuticals, and household cleaners. Carbopol®polymers may be used in either solid (e.g., powder) or liquid form.

The acrylic acid-based polymers suitable for use in the invention may behomopolymers or copolymers. Suitable homopolymers may be crosslinked,preferably with allyl sucrose or allylpentaerythritol. Suitablecopolymers of acrylic acid are modified by long chain (C₁₀-C₃₀) alkylacrylates and may be crosslinked, preferably with allylpentaerythritol.

Carbopol® polymers are neutralized in order to achieve maximumviscosity. As supplied, Carbopol® polymers are dry, tightly coiledacidic molecules, held in a coiled structure by hydrogen bonds. Oncedispersed in water, or another solvent, they begin to hydrate andpartially uncoil. The most common way to achieve maximum thickening fromCarbopol® polymers is by converting the acidic polymer into a salt. Thisis easily achieved by neutralizing with a common base such as sodiumhydroxide (NaOH) or triethanolamine (TEA). This neutralization “uncoils”the long chain polymer, swelling the molecule into an effectivethickening form.

Suitable thickening agents will yield the desired viscosity for theformulation, as well as other characteristics, such as appearance, shearresistance, ion resistance, and thermal stability. For example,Carbopol® 934 is preferred for a formulation that is either a suspensionor emulsion (rather than a clear gel) with a viscosity greater than 3000centipoise (cps). Carbopol® 974P may alternatively be used for itsadvantageous bioadhesive properties.

Any suitable amount of a thickening agent is present in the formulationof the invention to yield the desired viscosity for the formulation.Generally, the amount of thickening agent is from about 0.1% by weightto about 50% by weight, based on the weight of the formulation.Preferably, the amount of thickening agent is from about 0.1% to about10% by weight.

In other terms, the amount of thickening agent based on the volume ofthe ORP water solution is generally from about 0.1% weight/volume(mg/mL) to about 50% weight/volume (mg/mL). Preferably, the amount ofthickening agent is from about 0.1% w/v to about 10% w/v.

The amount of thickening agent generally is from about 0.1 g/250 mL toabout 50 mg/250 mL of the ORP water solution. Preferably, the amount ofthickening agent present is from about 1 mg/250 mL to about 20 mg/250 mLof the ORP water solution and, most preferably, from about 3 mg/250 mLto about 15 mg/250 mL.

When acrylic acid-based polymers are used at low concentrations, theformulation flows easily with a slippery feel. At higher concentrations,the formulation of the invention has a high viscosity and ispseudoplastic and resistant to flow. When shear force is applied by amixer or pump, the apparent viscosity is reduced, and the formulationmay be pumped.

The formulation of the invention may optionally include a neutralizingagent. Any suitable neutralizing agent may be used to yield the desiredpH of the formulation. Suitable neutralizing agents include, forexample, sodium hydroxide, triethanolamine, ammonia, potassiumhydroxide, L-arginine, AMP-95, Neutrol TE, Tris Amino, Ethomeen,di-isopropanolamine, and tri-isopropanolamine. Other neutralizing agentsare generally known in the art (see, e.g., Handbook of Cosmetic andPersonal Care Additives, 2nd ed., Ashe et al. eds. (2002), and Handbookof Pharmaceutical Excipients, 4th ed., Rowe et al. eds. (2003)).Suitable neutralizing agents may be either in liquid or solid form.

Preferably, the neutralizer triethanolamine used when the thickeningagent is an acrylic acid-based polymer such as Carbopol®. Theneutralizing agent converts the formulation into a gel.

Any suitable amount of neutralizing agent may be included in theformulation of the invention. Generally, the amount of neutralizingagent is from about 0.1% by weight to about 50% by weight, based on theweight of the formulation. Preferably, the amount of neutralizing agentis from about 0.1% to about 10% by weight, based on the weight of theformulation. On a volume basis, the amount of neutralizing agent ispresent in an amount of about 1% to about 50% by volume, based on thevolume of the ORP water solution.

When added in liquid form, the neutralizing may be added in an amount offrom about 1 mL/250 mL to about 100 mL/250 mL of the ORP water solution.Preferably, the amount of neutralizing agent is from about 10 mL/250 mLto about 90 mg/250 mL of the ORP water solution. Additionally, when insolid form, the neutralizing agent may be added in solid amounts whichcorrespond to these liquid amounts.

The formulation may further contain additional components such ascolorants, fragrances, buffers, physiologically acceptable carriersand/or excipients, and the like. Examples of suitable colorants include,but are not limited to, titanium dioxide, iron oxides, carbazole violet,chromium-cobalt-aluminum oxide,4-Bis[(2-hydroxyethyl)amino]-9,10-anthracenedione bis(2-propenoic)estercopolymers, and the like. Any suitable fragrance can be used.

The formulation of the invention may be prepared by any suitable means.The components of the formulation, such as the ORP water solution andthickening agent, may be mixed together in any manner to yield ahomogenous mixture. Preferably, the components are mixed together forseveral minutes using an electric mixture or other suitable device toensure uniformity. The components of the formulation are generally mixedfrom about 400 rpm to about 1000 rpm, preferably from about 500 rpm toabout 800 rpm, and more preferably from about 500 rpm to about 600 rpm.

The formulation is mixed for a sufficient period of time to yield ahomogenous mixture, generally from about 1 minute to about 10 minutesafter all of the components have been combined.

When the thickening agent is in the form of a power, it may first besieved to break up large agglomerates to allow for the preparation of ahomogenous formulation.

A neutralizing agent, such as triethanolamine, may subsequently be addedto the formulation containing the ORP water solution and thickeningagent. As noted above, the addition of triethanolamine may allow thethickening agent, such as Carbopol®, to uncoil and, thus, yield aformulation having the desired viscosity.

A colorant or fragrance may also be added to the mixture either beforeor after the thickening agent, such as Carbopol®, is dissolved into theORP water, but before the neutralization step.

The physical properties of the formulation of the invention aretypically the same as those of the ORP water solution present in theformulation. The properties of the ORP water solution remain even afterthe addition of a thickening agent and optional neutralizing agent. Forexample, the stability and pH of the ORP water solution itself and theformulation containing the ORP water solution are generally the same.Accordingly, all of the characteristics of the ORP water solutiondescribed herein apply to the formulation of the invention.

For example, the formulation of the invention is generally stable for atleast twenty-hours, and typically at least two days. More typically, theformulation is stable for at least about one week (e.g., one week, twoweeks, three weeks, four weeks, etc.), and preferably at least about twomonths. More preferably, the formulation is stable for at least sixmonths after its preparation. Even more preferably, the formulation isstable for at least one year, and most preferably for at least threeyears.

The pH of the formulation is generally from about 6 to about 8.Preferably, the pH of the formulation is from about 6.2 and about 7.8,and most preferably from about 7.4 and about 7.6.

The formulation of the invention may be used any form suitable fortopical administration to a patient. A suitable form includes, but isnot limited to, gel, lotion, cream, paste, ointment, and the like, whichforms are known in the art (see, e.g., Modern Pharmaceutics, 3rd ed.,Banker et al. ed. (1996)). Gels are typically a semisolid emulsion orsuspension that has a three-dimensional structure. Preferably, theformulation is in the form of a gel.

Pastes are generally semisolid suspensions that often contain a largeportion of solids (e.g., from about 20% to about 50%) dispersed in anaqueous or fatty vehicle. Lotions are typically liquid emulsionscontaining a water-based vehicle and volatiles (more than 50%) and thathave a sufficiently low viscosity (less than 30,000 cps) to be poured.Ointments and creams are generally semisolid emulsions or suspensionsthat may contain hydrocarbons or polyethylene glycols as part of thecarrier along with other volatile components.

When the formulation of the invention is in the form of a gel, theviscosity of the gel is in the range of from about 10,000 to about100,000 centipoise (cps) (e.g., about 15,000 cps, about 20,000 cps,about 25,000 cps, about 30,000 cps, about 35,000 cps, about 40,000 cps,about 45,000 cps, about 50,000 cps, about 55,000 cps, about 60,000 cps,about 65,000 cps, about 70,000 cps, about 75,000 cps, about 80,000 cps,about 85,000 cps, about 90,000 cps, about 95,000 cps, or rangesthereof), at about room temperature (e.g., about 25° C.).

The pH of the gel is typically from about 6.0 to about 8.0. Above thispH, the viscosity of the thickening agent, such as the Carbopol®polymer, may decrease leading to an unsatisfactory topical formulation.Preferably, the pH of the gel is from about 6.4 to about 7.8, and morepreferably, from about 7.4 to about 7.6.

The formulation of the invention is suitable for topical administrationto a patient, including a human and/or animal, to treat a variety ofconditions. Specifically, the formulation may be applied to animals(e.g., mice, rats, pigs, cows, horses, dogs, cats, rabbits, guinea pigs,hamsters, birds) and humans. Topical administration includes applicationto the skin as well as oral, intranasal, intrabronchial, and rectalroutes of administration.

In another embodiment, the invention is directed to a method fortreating a condition in a patient by topically administering aformulation comprising an ORP water solution and a thickening agent.

Conditions in a patient that may be treated according to the inventioninclude, for example, the following: surgical/open wound cleansingagent; skin pathogen disinfection (e.g., for bacteria, mycoplasmas,virus, fungi, prions); wound disinfection (e.g., battle wounds); woundhealing promotion; burn healing promotion; treatment of skin fungi;psoriasis; athlete's foot; ear infections (e.g., swimmer's ear);traumatic wounds; acute, subchronic and chronic infections (e.g.diabetic foot infections being an example of the latter), pressureulcers, derma-abrasion, debrided wounds, laser re-surfacing, donorsites/grafts, exuding partial and full thickness wounds, superficialinjuries (lacerations, cuts, abrasions, minor skin irritations) andother medical applications on or in the human or animal body. Ulcerstreated according to the invention may or may not have abscesses ornecrotic tissue present.

Additionally, the invention is directed to a method for promoting woundhealing in a patient by applying to a wound a formulation comprising anoxidative reductive potential water solution and a thickening agent. Thewound to be treated may be caused by any surgery, ulcer or other means.Ulcers that may be treated include, for example, diabetic foot ulcers.

The invention further relates to a method for preventing a condition ina patient by topically administering a formulation comprising an ORPwater solution and a thickening agent. For example, the formulation(e.g., in the form of a gel) can be used as a barrier on open wounds toprevent infection. Specifically, the formulation (e.g., in the form of agel) can be applied to the surface of a wound, such as a foot ulcerationin a diabetic, who is prone to neurological and vascular complications.The formulation applied thusly can provide a barrier to infection, sincethese wounds are the principal portal for infection for diabeticpatients.

The formulation may be used to prevent sexually transmitted diseases ina patient including, for example, infections. Such infections that maybe prevented include herpes, human immunodeficiency virus (HIV) andvaginal infections. When the formulation is in the form of a gel, it maybe used as a spermicide.

The formulation of the invention may be used or applied in atherapeutically effective amount to provide the desired therapeuticeffect on bacteria, viruses, and/or germs. As used herein, atherapeutically effective amount refers to an amount of the formulationthat results in an improvement of the condition being treated or to beprevented. For example, when used to treat an infection, atherapeutically effective amount of the formulation reduces the extentof the infection and/or prevents further infection. As is appreciated byone skilled in the art, the efficacy of the formulation of the inventionresulting from administering the formulation may be short-term (e.g., afew days) and/or long-term (e.g., months).

The formulation may further be applied over a sufficient period of time,for example, one two, several days, about one week, or several weeks,until the desired effect on the patient is observed.

The formulation may be applied in any suitable manner. For example, aquantity of the formulation may be applied to the surface of the patientto be treated and then evenly spread using the patient's own fingers.Alternatively, a health care provider may apply the formulation to thepatient's tissue. A suitable implement, for example, a disposable wipeor cloth, may be used to apply the formulation.

The ORP water of the present invention is produced by anoxidation-reduction process, which can be referred to as an electrolyticor redox reaction, in which electrical energy is used to producechemical change in an aqueous solution. Electrical energy is introducedinto and transported through water by the conduction of electricalcharge from one point to another in the form of an electrical current.In order for the electrical current to arise and subsist there must becharge carriers in the water, and there must be a force that makes thecarriers move. The charge carriers can be electrons, as in the case ofmetal and semiconductors, or they can be positive and negative ions inthe case of solutions.

A reduction reaction occurs at the cathode while an oxidation reactionoccurs at the anode in the process for preparing an ORP water solutionaccording to the invention. The specific reductive and oxidativereactions that occur are described in International Application WO03/048421 A1.

As used herein, water produced at an anode is referred to as anode waterand water produced at a cathode is referred to as cathode water. Anodewater contains oxidized species produced from the electrolytic reactionwhile cathode water contains reduced species from the reaction.

Anode water generally has a low pH typically of from about 1 to about6.8. Anode water generally contains chlorine in various forms including,for example, chlorine gas, chloride ions, hydrochloric acid and/orhypochlorous acid. Oxygen in various forms may also present including,optionally, e.g., oxygen gas, peroxides, and/or ozone. Cathode watergenerally has a high pH typically of from about 7.2 to about 11. Cathodewater generally contains hydrogen gas, hydroxyl radicals, and/or sodiumions.

The ORP water solution of the invention may be acidic, neutral or basic,and generally has a pH of from about 1 to about 14. At this pH, the ORPwater solution can safely be applied in suitable quantities to hardsurfaces without damaging the surfaces or harming objects, such as humanskin, that comes into contact with the ORP water solution. Typically,the pH of the ORP water solution is from about 3 to about 8. Morepreferably, the pH of the ORP water solution is from about 6.4 to about7.8, and most preferably, the pH is from about 7.4 to about 7.6.

The ORP water solution of the present invention generally has anoxidation-reduction potential of from about −1000 millivolts (mV) toabout +1350 millivolts (mV). This potential is a measure of the tendency(i.e., the potential) of a solution to either accept or transferelectrons that is sensed by a metal electrode and compared with areference electrode in the same solution. This potential may be measuredby standard techniques including, for example, by measuring theelectrical potential in millivolts of the ORP water solution relative tostandard reference silver/silver chloride electrode. The ORP watergenerally has a potential from about −400 mV to about +1300 mV or about+1150 mV. Preferably, the ORP water solution has a potential from about0 mV to about +1250 mV, and more preferably from about +500 mV to about+1250 mV. Even more preferably, the ORP water of the present inventionhas a potential of from about +800 mV to about +1100 mV, and mostpreferably from about +800 mV to about +1000 mV.

Various ionic and other species may be present in the ORP water solutionof the invention. For example, the ORP water solution may containchlorine (e.g., free chlorine and bound chlorine), and optionally, ozoneand peroxides (e.g., hydrogen peroxide). The presence of one or more ofthese species is believed to contribute to the disinfectant ability ofthe ORP water solution to kill a variety of microorganisms, such asbacteria and fungi, as well as viruses.

Free chlorine typically includes, but is not limited to, hypochlorousacid (HClO), hypochlorite ions (ClO⁻), sodium hypochlorite (NaOCl),chloride ion (Cl⁻), chlorite ions (ClO₂ ⁻), dissolved chlorine gas(Cl₂), and other radical chlorine species. The ratio of hypochlorousacid to hypochlorite ion is dependent upon pH. At a pH of 7.4,hypochlorous acid levels are from about 25 ppm to about 75 ppm.Temperature also impacts the ratio of the free chlorine component.

Bound chlorine is chlorine in chemical combination with ammonia ororganic amines (e.g., chloramines). Bound chlorine is generally presentin an amount up to about 20 ppm.

Chlorine and, optionally, ozone and hydrogen peroxide may present in theORP water solution of the invention in any suitable amount. The levelsof these components may be measured by methods known in the art.

Typically, the total chlorine content, which includes both free chlorineand bound chlorine, is from about 50 parts per million (ppm) to about200 ppm. Preferably, the total chlorine content is about 80 ppm to about150 ppm.

The chlorine content may be measured by methods known in the art, suchas the DPD colorimeter method (Lamotte Company, Chestertown, Md.) orother known methods established by the Environmental Protection Agency.In the DPD colorimeter method, a yellow color is formed by the reactionof free chlorine with N,N-diethyl-p-phenylenediamine (DPD) and theintensity is measured with a calibrated calorimeter that provides theoutput in parts per million. Further addition of potassium iodide turnsthe solution a pink color to provide the total chlorine value. Theamount of bound chlorine present is then determined by subtracting freechlorine from the total chlorine.

Ozone is optionally present in an amount of from about 0.03 ppm to about0.2 ppm, and preferably from about 0.10 ppm to about 0.16 ppm.

Hydrogen peroxide is optionally present at levels in the ORP watersolution in the range from about 0.01 ppm to about 200 ppm, andpreferably from about 0.05 ppm to about 100 ppm. More preferably,hydrogen peroxide is present in an amount from about 0.1 ppm and toabout 40 ppm, and most preferably from about 1 ppm to 4 ppm. Peroxidesare optionally present (e.g., H₂O₂, H₂O₂ ⁻ and HO₂ ⁻) in a concentrationof less than 0.12 milliMolar (mM).

The total amount of oxidizing chemical species present in the ORP watersolution is in the range of about 2 millimolar (mM) which includes theaforementioned chlorine species, oxygen species, and additional speciesthat may be difficult to measure such as Cl⁻, ClO₃, Cl₂ ⁻, and ClO_(x).The level of oxidizing chemical species present may also be measured byESR spectroscopy (using Tempone H as the spin trap molecule).

The ORP water solution of the invention is generally stable for at leastabout twenty-four hours, and typically at least about two days. Moretypically, the water solution is stable for at least about one week(e.g., one week, two weeks, three weeks, four weeks, etc.), andpreferably at least about two months. More preferably, the ORP watersolution is stable for at least about six months after its preparation.Even more preferably, the ORP water solution is stable for at leastabout one year, and most preferably for at least about three years.

As used herein, the term stable generally refers to the ability of theORP water solution remain suitable for its intended use, for example, indecontamination, disinfection, sterilization, anti-microbial cleansing,and wound cleansing, for a specified period of time after itspreparation under normal storage conditions (i.e., room temperature).

The ORP water solution of the invention is also stable when stored underaccelerated conditions, typically from about 30° C. to about 60° C., forat least about 90 days, and preferably about 180 days.

The concentrations of ionic and other species present solution aregenerally maintained during the shelf-life of the ORP water solution.Typically, the concentrations of free chlorine, and optionally, ozoneand hydrogen peroxides are maintained at about 70% or great from theirinitial concentration for at least about two months after preparation ofthe ORP water solution. Preferably, these concentrations are maintainedat about 80% or greater of their initial concentration for at leastabout two months after preparation of the ORP water solution. Morepreferably, these concentrations are at about 90% or greater of theirinitial concentration for at least about two months after preparation ofthe ORP water solution, and most preferably, about 95% or greater.

The stability of the ORP water solution of the invention may bedetermined based on the reduction in the amount of organisms present ina sample following exposure to the ORP water solution. The measurementof the reduction of organism concentration may be carried out using anysuitable organism including bacteria, fungi, yeasts, or viruses.Suitable organisms include, but are not limited to, Escherichia coli,Staphylococcus aureus, Candida albicans, and Bacillus athrophaeus(formerly B. subtilis). The ORP water solution is useful as both alow-level disinfectant capable of an about four log (10⁴) reduction inthe concentration of live microorganisms and a high-level disinfectantcapable of an about six log (10⁶) reduction in concentration of livemicroorganisms.

In one aspect of the invention, the ORP water solution is capable ofyielding at least about a four log (10⁴) reduction in total organismconcentration following exposure for one minute, when measured at leasttwo months after preparation of the solution. Preferably, the ORP watersolution is capable of such a reduction of organism concentration whenmeasured at least six months after preparation of the solution. Morepreferably, the ORP water solution is capable of such a reduction oforganism concentration when measured at least about one year afterpreparation of the ORP water solution, and most preferably when measuredat least about three years after preparation of the ORP water solution.

In another aspect of the invention, the ORP water solution is capable ofat least an about six log (10⁶) reduction in the concentration of asample of live microorganisms selected from the group consisting ofEscherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus andCandida albicans within one minute of exposure, when measured at leastabout two months after preparation of the ORP water solution.Preferably, the ORP water solution is capable of achieving thisreduction of Escherichia coli, Pseudomonas aeruginosa, Staphylococcusaureus or Candida albicans organisms when measured at least six monthsafter preparation, and more preferably at least one year afterpreparation. Preferably, the ORP water solution is capable of at leastan about seven log (10⁷) reduction in the concentration of such livemicroorganism within one minute of exposure, when measured at leastabout two months after preparation.

The ORP water solution of the invention is generally capable of reducinga sample of live microorganisms including, but not limited to,Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus andCandida albicans, from an initial concentration of from about 1×10⁶ toabout 1×10⁸ organisms/ml to a final concentration of about zeroorganisms/ml within one minute of exposure, when measured at least twomonths after preparation of the ORP water solution. This is between anabout six log (10⁶) to an about eight log (10⁸) reduction in organismconcentration. Preferably, the ORP water solution is capable ofachieving this reduction of Escherichia coli, Pseudomonas aeruginosa,Staphylococcus aureus or Candida albicans organisms when measured atleast about six months after preparation, and more preferably at leastabout one year after preparation.

Alternatively, the ORP water solution is capable of an about six log(10⁶) reduction in the concentration of a spore suspension of Bacillusathrophaeus spores within about five minutes of exposure, when measuredat least about two months after preparation of the ORP water solution.Preferably, the ORP water solution is capable of achieving thisreduction in the concentration of Bacillus athrophaeus spores whenmeasured at least about six months after preparation, and morepreferably at least about one year after preparation.

The ORP water solution is further capable of an about four log (10⁴)reduction in the concentration of a spore suspension of Bacillusathrophaeus spores within about thirty (30) seconds of exposure, whenmeasured at least about two months after preparation of the ORP watersolution. Preferably, the ORP water solution is capable of achievingthis reduction in the concentration of Bacillus athrophaeus spores whenmeasured at least about six months after preparation, and morepreferably at least about one year after preparation.

The ORP water solution is also capable of an about six log (10⁶)reduction in the concentration of fungal spores, such as Aspergillisniger spores, within about five to about ten minutes of exposure, whenmeasured at least about two months after preparation of the ORP watersolution. Preferably, the ORP water solution is capable of achievingthis reduction in the concentration of fungal spores when measured atleast about six months after preparation, and more preferably at leastabout one year after preparation.

In one embodiment, the ORP water solution of the invention optionallycomprises hydrogen peroxide (H₂O₂) and one or more chlorine species.Preferably, the chlorine species present is a free chlorine species. Thefree chlorine species may be selected from the group consisting ofhypochlorous acid (HOCl), hypochlorite ions (OCl⁻), sodium hypochlorite(NaOCl), chlorite ions (ClO₂ ⁻), chloride ion (Cl⁻), dissolved chlorinegas (Cl₂), and mixtures thereof.

Hydrogen peroxide is optionally present in the ORP water solutiongenerally in the range of from about 0.01 ppm to about 200 ppm, andpreferably from about 0.05 ppm to about 100 ppm. More preferably,hydrogen peroxide is present in an amount from about 0.1 ppm to about 40ppm, and most preferably from about 1 ppm to about 4 ppm.

The total amount of free chlorine species is generally from about 10 ppmto about 400 ppm, preferably from about 50 ppm to about 200 ppm, andmost preferably from about 50 ppm to about 80 ppm. The amount ofhypochlorous acid is in the generally from about 15 ppm to about 35 ppm.The amount of sodium hypochlorite is generally in the range of fromabout 25 ppm to about 50 ppm. Chlorine dioxide levels are generally lessthan about 5 ppm.

Generally, the ORP water solution is stable for at least about one week.Preferably, the ORP water solution is stable for at least about twomonths, More preferably, the ORP water solution is stable for at leastabout six months after its preparation. Even more preferably, the ORPwater solution is stable for at least about one year, and mostpreferably for at least about three years.

The pH of the ORP water solution in this embodiment is generally fromabout 6 to about 8. Preferably, the pH of the ORP water solution is fromabout 6.2 to about 7.8, and most preferably from about 7.4 to about 7.6.

While in no way limiting the present invention, it is believed that thecontrol of pH permits a stable ORP water solution in which chlorinespecies, such as, by way of example, hypochlorous acid and hypochloriteions, coexist.

Following its preparation, the ORP water solution or the formulation ofthe invention may be transferred to a sealed container for distributionand sale to end users such as, for example, health care facilitiesincluding hospitals, nursing homes, doctor offices, outpatient surgicalcenters, dental offices, and the like. The pharmaceutical dosage formaccording to the present invention comprises the formulation for topicaladministration as described herein and a sealed container into which theformulation is placed.

Any suitable sealed container may be used that maintains the sterilityand stability of the ORP water solution or formulation held by thecontainer. The container may be constructed of any material that iscompatible with the ORP water solution or the components of theformulation, for example, the ORP water solution and the thickeningagent. The container should be generally non-reactive so that the ionspresent in the ORP water solution do not react with the container to anyappreciable extent.

Preferably, the container is constructed of plastic or glass. Theplastic may be rigid so that the container is capable of being stored ona shelf. Alternatively, plastic may be flexible, such as a flexible bag.

Suitable plastics include polypropylene, polyester terephthalate (PET),polyolefin, cycloolefin, polycarbonate, ABS resin, polyethylene,polyvinyl chloride, and mixtures thereof. Preferably, the containercomprises polyethylene selected from the group consisting ofhigh-density polyethylene (HDPE), low-density polyethylene (LDPE), andlinear low-density polyethylene (LLDPE). Most preferably, the containeris high density polyethylene.

The container has an opening to permit dispensing of the ORP watersolution or formulation for administration to a patient. The containeropening may be sealed in any suitable manner. For example, the containermay be sealed with a twist-off cap or stopper. Optionally, the openingmay be further sealed with a foil layer.

The headspace gas of the sealed container may be air or other suitablegas that does not react with the ORP water solution or other componentsof a formulation containing the ORP water solution. Suitable headspacegases included nitrogen, oxygen, and mixtures thereof.

The invention further provides an ORP water solution comprising anodewater and cathode water. Anode water is produced in the anode chamber ofthe electrolysis cell used in the present invention. Cathode water isproduced in the cathode chamber of the electrolysis cell.

Cathode water is generally present in the ORP water solution of thesolution in an amount of from about 10% by volume to about 90% by volumeof the solution. Preferably, cathode water is present in the ORP watersolution in an amount of from about 10% by volume to about 50% byvolume, more preferably of from about 20% by volume to about 40% byvolume of the solution, and most preferably of from about 20% by volumeto about 30% by volume of the solution. Additionally, anode water may bepresent in the ORP water solution in an amount of from about 50% byvolume to about 90% by volume of the solution.

As noted herein, the ORP water solution containing both anode water andcathode water can be acidic, neutral or basic, and generally has a pH offrom about 1 to about 14. Typically, the pH of the ORP water solution isfrom about 3 to about 8. Preferably, the pH is from about 6.4 to about7.8, and more preferably from about 7.4 to about 7.6.

The ORP water solution of the invention has a wide variety of uses as adisinfectant, cleanser, cleaner, antiseptic and the like to control theactivity of unwanted or harmful substances present in the environment.Substances that may be treated with the ORP water solution include, forexample, organisms and allergens.

The ORP water solution may be used as a disinfectant, sterilizationagent, decontaminant, antiseptic and/or cleanser. The ORP water solutionof the invention is suitable for use in the following representativeapplications: medical, dental and/or veterinary equipment and devices;food industry (e.g., hard surfaces, fruits, vegetables, meats);hospitals/health care facilities (e.g., hard surfaces); cosmeticindustry (e.g., skin cleaner); households (e.g., floors, counters, hardsurfaces); electronics industry (e.g., cleaning circuitry, hard drives);and bio-terrorism (e.g., anthrax, infectious microbes).

The ORP water solution may also be applied to humans and/or animals totreat various conditions including, for example, the following:surgical/open wound cleansing agent; skin pathogen disinfection (e.g.,for bacteria, mycoplasmas, virus, fungi, prions); battle wounddisinfection; wound healing promotion; burn healing promotion; treatmentof stomach ulcers; wound irrigation; skin fungi; psoriasis; athlete'sfoot; pinkeye and other eye infections; ear infections (e.g., swimmer'sear); lung/nasal/sinus infections; and other medical applications on orin the human or animal body. The use of ORP water solutions as a tissuecell growth promoter is further described in U.S. Patent ApplicationPublication 2002/0160053 A1.

While in no way limiting the present invention, it is believed that theORP water solution eradicates the bacteria with which it contacts aswell as destroying the bacterial cellular components including proteinsand DNA.

For instance, the ORP water solution is capable of at least about fivelog (10⁵) reduction in the concentration of a sample of livemicroorganism selected from the group consisting of Pseudomonasaeruginosa, Escherichia coli, Enterococcus hirae, Acinetobacterbaumannii, Acinetobacter species, Bacteroides fragilis, Enterobacteraerogenes, Enterococcus faecalis, Vancomycin Resistant-Enterococcusfaecium (VRE, MDR), Haemophilus influenzae, Klebsiella oxytoca,Klebsiella pneumoniae, Micrococcus luteus, Proteus mirabilis, Serratiamarcescens, Staphylococcus aureus, Staphylococcus epidermidis,Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcussaprophyticus, Streptococcus pneumoniae, Streptococcus pyogenes, Candidaalbicans and Candida tropicalis, within 30 seconds of exposure, whenmeasured at least two months after preparation of the ORP watersolution.

In one embodiment, the ORP water solution administered in accordancewith the invention can reduce a sample of live microorganisms including,but not limited to, Escherichia coli, Pseudomonas aeruginosa,Staphylococcus aureus and Candida albicans, from an initialconcentration of from about 1×10⁶ to about 1×10⁸ organisms/ml to a finalconcentration of about zero organisms/ml within about one minute ofexposure when measured at least about two months after preparation ofthe ORP water solution. This corresponds to from about a six log (10⁶)to about an eight log (10⁸) reduction in organism concentration.Preferably, the ORP water solution is capable of achieving an about10⁶—about 10⁸ reduction of Escherichia coli, Pseudomonas aeruginosa,Staphylococcus aureus or Candida albicans organisms when measured atleast about six months after preparation, and more preferably whenmeasured at least about one year after preparation.

Alternatively, the ORP water solution administered in accordance withthe present invention can produce about a six log (10⁶) reduction in theconcentration of a spore suspension of Bacillus athrophaeus sporeswithin about five minutes of exposure when measured at least about twomonths after preparation of the ORP water solution. Preferably, the ORPwater solution administered in accordance with the invention can achieveabout a 10⁶ reduction in the concentration of Bacillus athrophaeusspores when measured at least about six months after preparation, andmore preferably when measured at least about one year after preparation.The ORP water solution is further capable of an about four log (10⁴)reduction in the concentration of a spore suspension of Bacillusathrophaeus spores within about thirty (30) seconds of exposure, whenmeasured at least two months after preparation of the ORP watersolution. Preferably, the ORP water solution is capable of achievingthis reduction in the concentration of Bacillus athrophaeus spores whenmeasured at least about six months after preparation, and morepreferably at least one about year after preparation.

The ORP water solution is also capable of an about six log (10⁶)reduction in the concentration of fungal spores, such as Aspergillisniger spores, within about five to about ten minutes of exposure, whenmeasured at least two months after preparation of the ORP watersolution. Preferably, the ORP water solution is capable of achievingthis reduction in the concentration of fungal spores when measured atleast six months after preparation, and more preferably at least oneyear after preparation.

The ORP water solution administered in accordance with the inventionfurther can produce more than 3 log (10³) reduction in the concentrationof viruses, such as Human Immunodeficiency Virus (HIV) and adenovirus,after from an about five to an about ten minutes exposure when measuredat least about two months after preparation of the ORP water solution.Preferably, the ORP water solution can achieve a >10³ reduction in theconcentration of viruses when measured at least about six months afterpreparation, and more preferably when measured at least about one yearafter preparation.

The ORP water solution administered in accordance with the inventionfurther can completely inhibit the growth of Mycobacterium bovis with anabout five minutes exposure when measured at least about two monthsafter preparation of the ORP water solution. Preferably, the ORP watersolution can achieve the total inhibition in the concentration ofMycobacteria when measured at least about six months after preparation,and more preferably when measured at least about one year afterpreparation.

Accordingly, organisms that can be controlled, reduced, killed oreradicated by treatment with the ORP water solution include, but are notlimited to, bacteria, fungi, yeasts, and viruses. Susceptible bacteriainclude, but are not limited to, Escherichia coli, Staphylococcusaureus, Bacillus athrophaeus, Streptococcus pyogenes, Salmonellacholeraesuis, Pseudomonas aeruginosa, Shingella dysenteriae, and othersusceptible bacteria. Fungi and yeasts that may be treated with the ORPwater solution include, for example, Candida albicans and Trichophytonmentagrophytes. The ORP water solution may also be applied to virusesincluding, for example, adenovirus, human immunodeficiency virus (HIV),rhinovirus, influenza (e.g., influenza A), hepatitis (e.g., hepatitisA), coronavirus (responsible for Severe Acute Respiratory Syndrome(SARS)), rotavirus, respiratory syncytial virus, herpes simplex virus,varicella zoster virus, rubella virus, and other susceptible viruses.

In a preferred embodiment, the ORP water solution of the invention maybe administered to treat patients with first, second or third degreeburns. Patients having a combination of burns, such as second and thirddegree burns, may also be treated with the ORP water solution. Firstdegree burns affect the epidermis, or skin surface. Second degree burnsaffect the epidermis and the underlying dermis. Third degree burnsaffect the epidermis, dermis and the hypodermis. More preferably, theORP water solution is administered to treat patients with second orthird degree . Burns that are suitable for treating according to theinvention are caused by various injuries, including, for example,contact with fire, boiling liquids (e.g., water, milk, etc.), orelectricity, and generally extend to from about 0% to about 69% of thepatient's tissue.

The ORP water solution may be administered to patients with burns in anysuitable manner. The ORP water solution may be administered topically byspraying, bathing, soaking, wiping or otherwise moistening the burn. TheORP water solution is administered in an amount sufficient to treat theburn. The ORP water solution is administered to a burn at least once aday and preferably more than once per day. More preferably, the ORPwater solution is administered to a burn three times per day.

The ORP water solution may be applied directly to the burn area, forexample, by pouring from a container or spraying from a reservoir. Theburn may be sprayed using any suitable device. Preferably, ahigh-pressure irrigation device is used to spray the ORP water solutionover the burn.

The burn may be soaked by submersing the burn either partially orcompletely in the ORP water solution. The burn may soak for any suitableperiod of time. Generally, the burn is soaked in the ORP water solutionfor at least about one minute. Preferably, the burn is soaked for fromabout 5 minutes to about 15 minutes.

Alternatively, the ORP water solution may be applied to the burn using asubstrate such as, for example, gauze, that has been saturated with ORPwater. Preferably, the ORP water solution is applied by multiple methodsincluding spraying and the burn is both sprayed and soaking.

The burn may optionally be dressed by applying a moist wound dressingsaturated with the ORP water solution. In addition to the moist wounddressing, the burn may optionally be dressed with dry gauze and anadhesive covering. Any suitable suave, cream, gel and/or ointment mayalso be applied to the burn surface after the administration of the ORPwater solution.

In one embodiment, a patient having a burn requiring treatment issubject to a washing procedure using the ORP water solution of theinvention. The ORP water solution is first sprayed on the burn using ahigh-pressure irrigation device. Next, the burn is soaked in the ORPwater solution for a suitable period of time. After soaking, the burn isthen sprayed with the ORP water solution again. The burn is then allowedto sit in a moistened state for at least about five minutes. Thisprocedure is carried out at least once a day on a patient's burn,preferably twice a day, and more preferably three times per day. In thisembodiment, the surface of the burn is preferably not dressed in betweenadministrations of the ORP water solution.

Prior to the administration of the ORP water solution, the burn ispreferably subject to debridement therapy to remove hyperkeratinized,necrotic, and otherwise unhealthy tissue down to healthy appearingtissue. In debriding the burn, the wound margins are excised to healthybleeding tissue. The burn may be cleaned of debris after debridement.The ORP water solution administered in accordance with the presentinvention also can be used as the irrigation solution for hydrosurgerydevices that are used to debride skin ulcers. Suitable hydrosurgerydevices can include, for example, the VersaJet devices sold in theUnited States by Smith and Nephew, Debritom in Europe by Medaxis, JetOxin the United States and Europe by DeRoyal or PulsaVac in Italy. It isbelieved that the ORP water solution can act synergistically with thedevice by reducing the microbial load in the wound and by avoiding theformation of infectious mists during the debridement procedure. Thus thedevice may be used to debride the burn with continuous irrigation,reduce the infection process and avoid the formation of infectious mistsin accordance with the present invention.

The ORP water solution administered in accordance with the presentinvention also can be used as the irrigation solution for negativepressure devices that are used to reduce edema and increase the bloodflow. Suitable negative pressure devices can include, e.g., one or morevacuum assisted wound closure devices such as, e.g., the V.A.C.® andV.A.C.® Instill™ devices sold in the United States by Kinetic Concepts,Inc. It is believed that the ORP water solution can act synergisticallywith the device by controlling the inflammatory-allergic process whilereducing the microbial load. Thus the device may be applied to the openburn wound with intermittent or continuous irrigation to treat orprevent tissue infection or necrosis in accordance with the presentinvention.

Optionally, several adjuvant therapies can also be utilized inaccordance with the invention including bioengineered skin (Apligraf,Organogenesis, Inc., Canton), acellular skin substitutes (Oasis WoundMatrix, Healthpoint), ultrasonic application of ORP water solutions, andlocal oxygen replacement or hyperbaric oxygen treatment (such as, e.g.,hyperbaric boots, the Vent-Ox System).

If necessary, administration of ORP water solution can be used incombination with skin grafts to promote healing of the burn.

The administration of ORP solution optionally be combined with theadministration of topical and/or systemic antibiotics. Suitableantibiotics can include, without limitation, penicillin, cephalosporinsor other β-lactams, macrolides (e.g., erythromycin,6-O-methylerythromycin, and azithromycin), fluoroquinolones,sulfonamides, tetracyclines, aminoglycosides, clindamycin, quinolones,metronidazole, vancomycin, chloramphenicol, antibacterially effectivederivatives thereof, and combinations thereof. Suitable anti-infectiveagents also can include antifungal agents such as, for example,amphotericin B, fluconazole, flucytosine, ketoconazole, miconazole,derivatives thereof, and combinations thereof. Suitableanti-inflammatory agents can include, e.g., one or moreanti-inflammatory drugs, e.g., one or more anti-inflammatory steroids orone or more non-steroidal anti-inflammatory drugs (NSAIDs). Exemplaryanti-inflammatory drugs can include, e.g., cyclophilins, FK bindingproteins, anti-cytokine antibodies (e.g. anti-TNF), steroids, andNSAIDs.

In another embodiment of the invention, a second and/or third degreeburn on a patient is initially debrided and then sprayed with the ORPwater solution with a high-pressure irrigation device. The amount of ORPwater solution used to wash the burn is preferably sufficient to removedebris. The burn is then soaked in the ORP water solution for a suitableperiod of time. The patient's burn is next sprayed with ORP watersolution, and the solution is allowed to moisten the burn for a suitableperiod of time, preferably from about 5 minutes to about 15 minutes. Thespraying and moistening is repeated about three times a day. In betweenthe administrations of ORP water solution, the surface of the burn isnot dressed.

The process of high-pressure spraying, optionally soaking, spraying, andmoistening the burn may be repeated at suitable intervals. Preferably,the procedure in which the burn is high-pressure sprayed, optionallysoaked, sprayed, and moistened is repeated about once per week and morepreferably, about once per day. The treatment of the burn using the ORPwater solution may continue until the burn is sufficiently healed whichtypically requires repeating the procedure over several days. Generally,the ORP water solution is applied every day for at least about threedays. Typically, the ORP water solution is applied every day for atleast about five days, preferably for at least about seven days, andmore preferably for at least about ten days. The healing of the burn istypically measured by the rate of scar contraction and epithielization.

The ORP water of the invention is also suitable for use in controllingthe activity of allergens present in the environment. As used herein,allergens include any substance other than bacteria, fungi, yeasts, orviruses that can trigger an adverse immune response, or allergy, insusceptible people or animals. Asthma is a common physiological responsefollowing exposure to one or more allergens. Allergens may be eitherviable (i.e., from living or dead organisms) or non-viable (e.g.,non-living such as textiles), and may be present in the environment, forexample, in households and/or workplaces.

Protein-based household allergens that may be treated with the ORP waterinclude, for example, animal fur, skin, and feces, household dust,weeds, grasses, trees, mites, and pollens. Animal allergens include, forexample, cat epithelium, dog epithelium, horse dander, cow dander, dogdander, guinea pig epithelium, goose feathers, mouse epithelium, mouseurine, rat epithelium and rat urine.

Occupational allergens include, for example, high-molecular-weightagents, such as natural proteins generally derived from plant or animalproteins, and low-molecular-weight chemicals, such as diisocyanates, andother material found in some textiles. Other chemical allergens that maybe present in the workplace include, for example, anhydrides,antibiotics, wood dust and dyes. Numerous proteins may be occupationalallergens including vegetable gums, enzymes, animal proteins, insects,plant proteins, and legumes.

Additional allergens suitable for treatment by the ORP water solutionare described in Korenblat and Wedner, Allergy Theory and Practice(1992) and Middleton, Jr., Allergy Principles and Practice (1993).

It has been found that the ORP water solution administered in accordancewith the invention is virtually free of toxicity to normal tissues andnormal mammalian cells. The ORP water solution administered inaccordance with the invention causes no significant decrease in theviability of eukaryotic cells, no significant increase in apoptosis, nosignificant acceleration of cell aging and/or no significant oxidativeDNA damage in mammalian cells. The non-toxicity is particularlyadvantageous, and perhaps even surprising, given that the disinfectingpower of the ORP water solution administered in accordance with theinvention is roughly equivalent to that of hydrogen peroxide, yet issignificantly less toxic than hydrogen peroxide is to normal tissues andnormal mammalian cells. These findings demonstrate that the ORP watersolution administered in accordance with the present invention is safefor use, e.g., in mammals, including humans.

For the ORP water solution administered in accordance with theinvention, the cell viability rate is preferably at least about 65%,more preferably at least about 70%, and still more preferably at leastabout 75% after an about 30 minute exposure to the ORP water solution.In addition, the ORP water solution administered in accordance with theinvention preferably causes only up to about 10% of cells, morepreferably only up to about 5% of cells, and still more preferably onlyup to about 3% of cells to expose Annexin-V on their cellular surfaceswhen contacted with the ORP water solution for up to about thirtyminutes or less (e.g., after about thirty minutes or after about fiveminutes of contact with the ORP water solution). Further, the ORP watersolution administered in accordance with the invention preferably causesless than about 15% of cells, more preferably less than about 10% ofcells, and still more preferably less than about 5% of cells to expressthe SA-β-galactosidase enzyme after chronic exposure to the OPR watersolution. The ORP water solution administered in accordance with theinvention preferably causes caused the same fraction of the oxidativeDNA adduct formation caused by saline solution, e.g., less than about20% of the oxidative DNA adduct formation, less than about 10% of theoxidative DNA adduct formation, or about 5% or less of the oxidative DNAadduct formation normally caused by hydrogen peroxide in cells treatedunder equivalent conditions.

The ORP water solution administered in accordance with the inventionproduces no significant RNA degradation. Accordingly, RNA extracted fromhuman cell cultures after an about 30 minutes exposure to the ORP watersolution or r at about 3 hours after an about 30 minute-exposure, andanalyzed by denaturing gel electrophoresis, will typically show nosignificant RNA degradation and will typically exhibit two discreetbands corresponding to the ribosomal eukaryotic RNAs (i.e. 28S and 18S)indicating that the ORP water solution administered in accordance withthe invention leaves the RNA substantially intact. Similarly, RNAextracted from human cell cultures after about 30 minutes of exposure tothe ORP water solution or after about 3 hours of exposure, can besubjected reverse transcription and amplification (RT-PCR) of theconstitutive human GAPDH (Glyceraldehyde-3-phosphate dehydrogenase) geneand result in a strong GAPDH band on gel electrophoresis of the RT-PCRproducts. By contrast, cells treated with HP for a similar period showsignificant RNA degradation and little if any GAPDH RT-PCR product.

The ORP water solution of the invention may be used or applied in anysuitable amount to provide the desired bactericidal, virucidal,germicidal and/or anti-allergenic effect.

The ORP water solution may be applied to disinfect and sterilize in anysuitable manner. For example, to disinfect and sterilize medical ordental equipment, the equipment is maintained in contact with the ORPwater solution for a sufficient period of time to reduce the level oforganisms present on the equipment to a desired level.

For disinfection and sterilization of hard surfaces, the ORP watersolution may be applied to the hard surface directly from a container inwhich the ORP water solution is stored. For example, the ORP watersolution may be poured, sprayed or otherwise directly applied to thehard surface. The ORP water solution may then be distributed over thehard surface using a suitable substrate such as, for example, cloth,fabric or paper towel. In hospital applications, the substrate ispreferably sterile. Alternatively, the ORP water solution may first beapplied to a substrate such as cloth, fabric or paper towel. The wettedsubstrate is then contacted with the hard surface. Alternatively, theORP water solution may be applied to hard surfaces by dispersing thesolution into the air as described herein. The ORP water solution may beapplied in a similar manner to humans and animals.

The invention further provides a cleaning wipe comprising a waterinsoluble substrate and the ORP water solution as described herein,wherein the ORP water solution is dispensed onto the substrate. The ORPwater solution may be impregnated, coated, covered or otherwise appliedto the substrate. Preferably, the substrate is pretreated with the ORPwater solution before distribution of the cleaning wipes to end users.

The substrate for the cleaning wipe may be any suitable water-insolubleabsorbent or adsorbent material. A wide variety of materials can be usedas the substrate. It should have sufficient wet strength, abrasivity,loft and porosity. Further, the substrate must not adversely impact thestability of the ORP water solution. Examples include non wovensubstrates, woven substrates, hydroentangled substrates and sponges.

The substrate may have one or more layers. Each layer may have the sameor different textures and abrasiveness. Differing textures can resultfrom the use of different combinations of materials or from the use ofdifferent manufacturing processes or a combination thereof. Thesubstrate should not dissolve or break apart in water. The substrateprovides the vehicle for delivering the ORP water solution to thesurface to be treated.

The substrate may be a single nonwoven sheet or multiple nonwovensheets. The nonwoven sheet may be made of wood pulp, synthetic fibers,natural fibers, and blends thereof. Suitable synthetic fibers for use inthe substrate include, without limitation, polyester, rayon, nylon,polypropylene, polyethylene, other cellulose polymers, and mixtures ofsuch fibers. The nonwovens may include nonwoven fibrous sheet materialswhich include meltblown, coform, air-laid, spun bond, wet laid,bonded-carded web materials, hydroentangled (also known as spunlaced)materials, and combinations thereof. These materials can comprisesynthetic or natural fibers or combinations thereof. A binder mayoptionally be present in the substrate.

Examples of suitable nonwoven, water insoluble substrates include 100%cellulose Wadding Grade 1804 from Little Rapids Corporation, 100%polypropylene needlepunch material NB 701-2.8-W/R from AmericanNon-wovens Corporation, a blend of cellulosic and syntheticfibres-Hydraspun 8579 from Ahlstrom Fibre Composites, and 70%Viscose/30% PES Code 9881 from PGI Nonwovens Polymer Corp. Additionalexamples of nonwoven substrates suitable for use in the cleaning wipesare described in U.S. Pat. Nos. 4,781,974, 4,615,937, 4,666,621, and5,908,707, and International Patent Application Publications WO98/03713, WO 97/40814, and WO 96/14835.

The substrate may also be made of woven materials, such as cottonfibers, cotton/nylon blends, or other textiles. Regenerated cellulose,polyurethane foams, and the like, which are used in making sponges, mayalso be suitable for use.

The liquid loading capacity of the substrate should be at least about50%-1000% of the dry weight thereof, most preferably at least about200%-800%. This is expressed as loading ½ to 10 times the weight of thesubstrate. The weight of the substrate varies without limitation fromabout 0.01 to about 1,000 grams per square meter, most preferably 25 to120 grams/m2 (referred to as “basis weight”) and typically is producedas a sheet or web which is cut, die-cut, or otherwise sized into theappropriate shape and size. The cleaning wipes will preferably have acertain wet tensile strength which is without limitation about 25 toabout 250 Newtons/m, more preferably about 75-170 Newtons/m.

The ORP water solution may be dispensed, impregnated, coated, covered orotherwise applied to the substrate by any suitable method. For example,individual portions of substrate may be treated with a discrete amountof the ORP water solution. Preferably, a mass treatment of a continuousweb of substrate material with the ORP water solution is carried out.The entire web of substrate material may be soaked in the ORP watersolution. Alternatively, as the substrate web is spooled, or even duringcreation of a nonwoven substrate, the ORP water solution is sprayed ormetered onto the web. A stack of individually cut and sized portions ofsubstrate may be impregnated or coated with the ORP water solution inits container by the manufacturer.

The cleaning wipes may optionally contain additional components toimprove the properties of the wipes. For example, the cleaning wipes mayfurther comprise polymers, surfactants, polysaccharides,polycarboxylates, polyvinyl alcohols, solvents, chelating agents,buffers, thickeners, dyes, colorants, fragrances, and mixtures thereofto improve the properties of the wipes. These optional components shouldnot adversely impact the stability of the ORP water solution. Examplesof various components that may optionally be included in the cleaningwipes are described in U.S. Pat. Nos. 6,340,663, 6,649,584 and6,624,135.

The cleaning wipes of the invention can be individually sealed with aheat-sealable or glueable thermoplastic overwrap (such as polyethylene,Mylar, and the like). The wipes can also be packaged as numerous,individual sheets for more economical dispensing. The cleaning wipes maybe prepared by first placing multiple sheets of the substrate in adispenser and then contacting the substrate sheets with the ORP watersolution of the invention. Alternatively, the cleaning wipes can beformed as a continuous web by applying the ORP water solution to thesubstrate during the manufacturing process and then loading the wettedsubstrate into a dispenser.

The dispenser includes, but is not limited to, a canister with aclosure, or a tub with closure. The closure on the dispenser is to sealthe moist wipes from the external environment and to prevent prematurevolatilization of the liquid ingredients.

The dispenser may be made of any suitable material that is compatiblewith both the substrate and the ORP water solution. For example, thedispenser may be made of plastic, such as high density polyethylene,polypropylene, polycarbonate, polyethylene terephthalate (PET),polyvinyl chloride (PVC), or other rigid plastics.

The continuous web of wipes may be threaded through a thin opening inthe top of the dispenser, most preferably, through the closure. A meansof sizing the desired length or size of the wipe from the web would thenbe needed. A knife blade, serrated edge, or other means of cutting theweb to desired size may be provided on the top of the dispenser, fornon-limiting example, with the thin opening actually doubling in duty asa cutting edge. Alternatively, the continuous web of wipes may bescored, folded, segmented, perforated or partially cut into uniform ornon-uniform sizes or lengths, which would then obviate the need for asharp cutting edge. Further, the wipes may be interleaved, so that theremoval of one wipe advances the next.

The ORP water solution of the invention may alternatively be dispersedinto the environment through a gaseous medium, such as air. The ORPwater solution may be dispersed into the air by any suitable means. Forexample, the ORP water solution may be formed into droplets of anysuitable size and dispersed into a room.

For small scale applications, the ORP water solution may be dispensedthrough a spray bottle that includes a standpipe and pump.Alternatively, the ORP water solution may be packaged in aerosolcontainers. Aerosol containers generally include the product to bedispensed, propellant, container, and valve. The valve includes both anactuator and dip tube. The contents of the container are dispensed bypressing down on the actuator. The various components of the aerosolcontainer are compatible with the ORP water solution. Suitablepropellants may include a liquefied halocarbon, hydrocarbon, orhalocarbon-hydrocarbon blend, or a compressed gas such as carbondioxide, nitrogen, or nitrous oxide. Aerosol systems typically yielddroplets that range in size from about 0.15 μm to about 5 μm.

The ORP water solution may be dispensed in aerosol form as part of aninhaler system for treatment of infections in the lungs and/or airpassages or for the healing of wounds in such parts of the body.

For larger scale applications, any suitable device may be used todisperse the ORP water solution into the air including, but not limitedto, humidifiers, misters, foggers, vaporizers, atomizers, water sprays,and other spray devices. Such devices permit the dispensing of the ORPwater solution on a continuous basis. An ejector which directly mixesair and water in a nozzle may be employed. The ORP water solution may beconverted to steam, such as low pressure steam, and released into theair stream. Various types of humidifiers may be used such as ultrasonichumidifiers, stream humidifiers or vaporizers, and evaporativehumidifiers.

The particular device used to disperse the ORP water solution may beincorporated into a ventilation system to provide for widespreadapplication of the ORP water solution throughout an entire house orhealthcare facility (e.g., hospital, nursing home, etc.).

The present invention further provides a process for producing an ORPwater solution using at least one electrolysis cell comprising an anodechamber, cathode chamber and salt solution chamber located between theanode and cathode chambers, wherein the ORP water solution comprisesanode water and cathode water. A diagram of a typical three chamberelectrolysis cell useful in the invention is shown in FIG. 1.

The electrolysis cell 100 has an anode chamber 102, cathode chamber 104and salt solution chamber 106. The salt solution chamber is locatedbetween the anode chamber 102 and cathode chamber 104. The anode chamber102 has an inlet 108 and outlet 10 to permit the flow of water throughthe anode chamber 102. The cathode chamber 104 similarly has an inlet112 and outlet 114 to permit the flow of water through the cathodechamber 104. The salt solution chamber 106 has an inlet 116 and outlet118. The electrolysis cell 100 preferably includes a housing to hold allof the components together.

The anode chamber 102 is separated from the salt solution chamber by ananode electrode 120 and an anion ion exchange membrane 122. The anodeelectrode 120 may be positioned adjacent to the anode chamber 102 withthe membrane 122 located between the anode electrode 120 and the saltsolution chamber 106. Alternatively, the membrane 122 may be positionedadjacent to the anode chamber 102 with the anode electrode 120 locatedbetween the membrane 122 and the salt solution chamber 106.

The cathode chamber 104 is separated from the salt solution chamber by acathode electrode 124 and a cathode ion exchange membrane 126. Thecathode electrode 124 may be positioned adjacent to the cathode chamber104 with the membrane 126 located between the cathode electrode 124 andthe salt solution chamber 106. Alternatively, the membrane 126 may bepositioned adjacent to the cathode chamber 104 with the cathodeelectrode 124 located between the membrane 126 and the salt solutionchamber 106.

The electrodes are generally constructed of metal to permit a voltagepotential to be applied between the anode chamber and cathode chamber.The metal electrodes are generally planar and have similar dimensionsand cross-sectional surface area to that of the ion exchange membranes.The electrodes are configured to expose a substantial portion of thesurface of the ion exchange members to the water in their respectiveanode chamber and cathode chamber. This permits the migration of ionicspecies between the salt solution chamber, anode chamber and cathodechamber. Preferably, the electrodes have a plurality of passages orapertures evenly spaced across the surface of the electrodes.

A source of electrical potential is connected to the anode electrode 120and cathode electrode 124 so as to induce an oxidation reaction in theanode chamber 102 and a reduction reaction in the cathode chamber 104.

The ion exchange membranes 122 and 126 used in the electrolysis cell 100may be constructed of any suitable material to permit the exchange ofions between the salt solution chamber 106 and the anode chamber 102such as chloride ions (Cl−) and between the salt solution salt solutionchamber 106 and the cathode chamber 104 such as sodium ions (Na⁺). Theanode ion exchange membrane 122 and cathode ion exchange membrane 126may be made of the same or different material of construction.Preferably, the anode ion exchange membrane comprises a fluorinatedpolymer. Suitable fluorinated polymers include, for example,perfluorosulfonic acid polymers and copolymers such as perfluorosulfonicacid/PTFE copolymers and perfluorosulfonic acid/TFE copolymers. The ionexchange membrane may be constructed of a single layer of material ormultiple layers.

The source of the water for the anode chamber 102 and cathode chamber104 of the electrolysis cell 100 may be any suitable water supply. Thewater may be from a municipal water supply or alternatively pretreatedprior to use in the electrolysis cell. Preferably, the pretreated wateris selected from the group consisting of softened water, purified water,distilled water, and deionized water. More preferably, the pretreatedwater source is ultrapure water obtained using reverse osmosispurification equipment.

The salt water solution for use in the salt solution chamber 106 may beany aqueous salt solution that contains suitable ionic species toproduce the ORP water solution. Preferably, the salt water solution isan aqueous sodium chloride (NaCl) salt solution, also commonly referredto as a saline solution. Other suitable salt solutions include otherchloride salts such as potassium chloride, ammonium chloride andmagnesium chloride as well as other halogen salts such as potassium andbromine salts. The salt solution may contain a mixture of salts.

The salt solution may have any suitable concentration. The salt solutionmay be saturated or concentrated. Preferably, the salt solution is asaturated sodium chloride solution.

The various ionic species produced in the three chambered electrolysiscell useful in the invention are illustrated in FIG. 2. The threechambered electrolysis cell 200 includes an anode chamber 202, cathodechamber 204, and a salt solution chamber 206. Upon application of asuitable electrical current to the anode 208 and cathode 210, the ionspresent in the salt solution flowing through the salt solution chamber206 migrate through the anode ion exchange membrane 212 and cathode ionexchange membrane 214 into the water flowing through the anode chamber202 and cathode chamber 204, respectively.

Positive ions migrate from the salt solution 216 flowing through thesalt solution chamber 206 to the cathode water 218 flowing through thecathode chamber 204. Negative ions migrate from the salt solution 216flowing through the salt solution chamber 206 to the anode water 220flowing through the anode chamber 202.

Preferably, the salt solution 216 is aqueous sodium chloride (NaCl) thatcontains both sodium ions (Na+) and chloride ions (Cl−) ions. PositiveNa+ ions migrate from the salt solution 216 to the cathode water 218.Negative Cl− ions migrate from the salt solution 216 to the anode water220.

The sodium ions and chloride ions may undergo further reaction in theanode chamber 202 and cathode chamber 204. For example, chloride ionscan react with various oxygen ions and other species (e.g., oxygen freeradicals, O2, O3) present in the anode water 220 to produce ClOn- andClO—. Other reactions may also take place in the anode chamber 202including the formation of oxygen free radicals, hydrogen ions (H+),oxygen (as O2), ozone (O3), and peroxides. In the cathode chamber 204,hydrogen gas (H2), sodium hydroxide (NaOH), hydroxide ions (OH—),ClOn-ions, and other radicals may be formed.

The invention further provides for a process and apparatus for producingan ORP water solution using at least two three chambered electrolysiscells. A diagram of a process for producing an ORP water solution usingtwo electrolysis cells of the invention is shown in FIG. 3.

The process 300 includes two three-chambered electrolytic cells,specifically a first electrolytic cell 302 and second electrolytic cell304. Water is transferred, pumped or otherwise dispensed from the watersource 305 to anode chamber 306 and cathode chamber 308 of the firstelectrolytic cell 302 and to anode chamber 310 and cathode chamber 312of the second electrolytic cell 304. Typically, the process of theinvention can produce from about 1 liter/minute to about 50liters/minute of ORP water solution. The production capacity may beincreased by using additional electrolytic cells. For example, three,four, five, six, seven, eight, nine, ten or more three-chamberedelectrolytic cells may be used to in increase the output of the ORPwater solution of the invention.

The anode water produced in the anode chamber 306 and anode chamber 310is collected are collected in the mixing tank 314. A portion of thecathode water produced in the cathode chamber 308 and cathode chamber312 is collected in mixing tank 314 and combined with the anode water.The remaining portion of cathode water produced in the process isdiscarded. The cathode water may optionally be subjected to gasseparator 316 and/or gas separator 318 prior to addition to the mixingtank 314. The gas separators remove gases such as hydrogen gas that areformed in cathode water during the production process.

The mixing tank 314 may optionally be connected to a recirculation pump315 to permit homogenous mixing of the anode water and portion ofcathode water from electrolysis cells 302 and 304. Further, the mixingtank 314 may optionally include suitable devices for monitoring thelevel and pH of the ORP water solution. The ORP water solution may betransferred from the mixing tank 314 via pump 317 for application indisinfection or sterilization at or near the location of the mixingtank. Alternatively, the ORP water solution may be dispensed intosuitable containers for shipment to a remote site (e.g., warehouse,hospital, etc.).

The process 300 further includes a salt solution recirculation system toprovide the salt solution to salt solution chamber 322 of the firstelectrolytic cell 302 and the salt solution chamber 324 of the secondelectrolytic cell 304. The salt solution is prepared in the salt tank320. The salt solution is transferred via pump 321 to the salt solutionchambers 322 and 324. Preferably, the salt solution flows in seriesthrough salt solution chamber 322 first followed by salt solutionchamber 324. Alternatively, the salt solution may be pumped to both saltsolution chambers simultaneously.

Before returning to the salt tank 320, the salt solution may flowthrough a heat exchanger 326 in the mixing tank 314 to control thetemperature of the ORP water solution as needed.

The ions present in the salt solution are depleted over time in thefirst electrolytic cell 302 and second electrolytic cell 304. Anadditional source of ions may periodically be added to the mixing tank320 to replace the ions that are transferred to the anode water andcathode water. The additional source of ions may be used to maintain aconstant pH of the salt solution which tends to drop (i.e., becomeacidic) over time. The source of additional ions may be any suitablecompound including, for example, salts such as sodium chloride.Preferably, sodium hydroxide is added to the mixing tank 320 to replacethe sodium ions (Na⁺) that are transferred to the anode water andcathode water.

In another embodiment, the invention provides an apparatus for producingan oxidative reductive potential water solution comprising at least twothree-chambered electrolytic cells. Each of the electrolytic cellsincludes an anode chamber, cathode chamber, and salt solution chamberseparating the anode and cathode chambers. The apparatus includes amixing tank for collecting the anode water produced by the electrolyticcells and a portion of the cathode water produced by one or more of theelectrolytic cells. Preferably, the apparatus further includes a saltrecirculation system to permit recycling of the salt solution suppliedto the salt solution chambers of the electrolytic cells.

The following examples further illustrate the invention but, of course,should not be construed as in any way limiting in its scope.

EXAMPLES 1-3

These examples demonstrate the unique features of the ORP water solutionof the invention. The samples of the ORP water solution in Examples 1-3were analyzed in accordance with the methods described herein todetermine the physical properties and levels of ionic and other chemicalspecies present in each sample. The results obtained for chlorinedioxide, ozone and hydrogen peroxide are based on standard tests used tomeasure such species; however, the results may be indicative ofdifferent species, which can also generate positive test results.Further, it has been reported that chlorine dioxide, ozone and hydrogenperoxide can react with hypochlorite resulting in their consumption andthe production of other species (e.g., HCl and O₂). The pH,oxidative-reductive potential (ORP) and ionic species present are setforth in Table 1 for each sample of the ORP water solution.

TABLE 1 Physical Characteristics and Ion Species Present for the ORPWater Solution Sample EXAMPLE 1 EXAMPLE 2 EXAMPLE 3 pH 7.45 7.44 7.45ORP (mV) +879 +881 +874 Total Cl⁻ (ppm) 110 110 120 Bound Cl⁻ (ppm) 5 66

The ORP water solution has suitable physical characteristics for use indisinfection, sterilization and/or cleaning.

EXAMPLES 4-10

These examples demonstrate the addition of a bleaching agent to the ORPwater solution according to the invention in various amounts. Inparticular, these examples demonstrate the antimicrobial activity andfabric bleaching ability of the compositions.

A 10% Clorox® bleach solution was prepared using distilled water. Thefollowing solutions were then prepared using the 10% bleach solution:80% ORP water solution/20% bleach (Example 4); 60% ORP watersolution/40% bleach (Example 5); 40% ORP water solution/60% bleach(Example 6); 20% ORP water solution/80% bleach (Example 7); and 0% ORPwater solution/100% bleach (Example 8). Two control solutions were alsoused for comparison including 100% ORP water solution/0% bleach (Example9) and an ORP water solution with 0.01% Tween 20 detergent (Example 10).The physical characteristics of these samples were determined,specifically pH, oxidative-reductive potential (ORP), total chlorine(Cl⁻) content, hypochlorous acid (HClO⁻) content, chlorine dioxidecontent and peroxide content, and are set forth in Table 2.

TABLE 2 Physical Characteristics of ORP Water Solution/BleachCompositions ORP Total Cl⁻ HClO⁻ pH (mV) (ppm) (ppm) Ex. 4 8.92 +7891248 62 Ex. 5 9.20 +782 2610 104 Ex. 6 9.69 +743 4006 80 Ex. 7 9.86 +7304800 48 Ex. 8 9.80 +737 5000 50 Ex. 9 7.06 +901 64 32 Ex. 10 6.86 +91451 26

The large bolus of chlorine ions added as part of the bleaching agentprevented the accurate measurement of the chlorine dioxide and peroxidelevels as indicated with the n.d. designations. Also, the resultsobtained for chlorine dioxide and peroxide are based on standard testsused to measure such species; however, the results may be indicative ofdifferent species which can also generate positive test results.Further, it has been reported that chlorine dioxide, ozone and hydrogenperoxide can react with hypochlorite resulting their consumption and theproduction of other species (e.g., HCl and O₂). As these examplesdemonstrate, the hypochlorous acid levels of the ORP water solution withand without the addition of a bleaching agent are similar.

The samples of Examples 4-10 were subjected to a high spore count testusing Bacillus subtilis var. niger spores (ATCC #9372 obtained from SPSMedical of Rush, N.Y.). Spore suspensions were concentrated (byevaporation in a sterile hood) to 4×10⁶ spores per 100 microliters. A100 microliter sample of the spore suspension were mixed with 900microliters of each of the samples in Examples 4-10. The samples wereincubated at room temperature for periods of 1 to 5 minutes as set forthin Table 3. At the indicated times, 100 microliters of the incubatedsamples were plated onto individual TSA plates and incubated for 24hours at 35° C.+2° C., after which the number of resulting colonies oneach plate was determined. The control plates demonstrated that thestarting spore concentrations were >1×10⁶ spores/100 microliters. Theconcentration of Bacillus spores for the various samples at the variousincubation times (as the average of two determinations) is set forth inTable 3.

TABLE 3 Bacillus Spore Concentrations (spores/100 microliters) 1 minute2 minutes 3 minutes 4 minutes 5 minutes Ex. 4 >>1000 411 1 0 2 Ex.5 >>1000 1000 1 0 0 Ex. 6 >>1000 >>1000 >1000 22 0 Ex.7 >>1000 >>1000 >1000 15 0 Ex. 8 >>1000 >>1000 >1000 3 1 Ex. 9 >>1000 740 0 0 Ex 10 >>1000 239 3 0 0

As these results demonstrate, as the concentration of bleach (as 10%aqueous bleach solution) increases, the amount of Bacillus spores killedis reduced for the samples incubated for 2-3 minutes. However, forsamples incubated for 5 minutes, the bleach concentration does notimpact Bacillus spore kill. Further, the results demonstrate that theaddition of 0.01% detergent to the ORP water solution does not reducespore kill.

The samples of Examples 4-10 were subjected to a fabric bleaching test.The fabric upon which the samples were tested was a 100% rayonchildren's t-shirt with dark blue dye patches. Two inch square pieces ofdyed fabric were placed into 50 mL plastic tubes. Each fabric piece wascovered by a sample of the solution in Examples 4-10. The elapsed timeuntil complete bleaching was obtained, as determined by the whitening ofthe fabric, is set forth in Table 4.

TABLE 4 Time Until Complete Bleaching of Fabric Sample Example Time Ex.4 39 minutes Ex. 5 23 minutes Ex. 6 18 minutes Ex. 7 19 minutes Ex. 8 10minutes Ex. 9 >6 hours Ex. 10 >6 hours

As demonstrated by these examples, as the concentration of the ORP watersolution increases in the composition, the time until complete bleachingis achieved increases.

EXAMPLE 11

This example relates to the toxicological profile of on an ORP watersolution of the present invention. Microcyn 60 (or M60), an exemplaryORP water solution of the present invention, was used in these studies.

In terms of safety, M60 was not an irritant to the skin or conjuctiva ofrabbits as tested in compliance with international standards (AAMI 1997;NV SOP 16G-44; PFEUM 2000). Furthermore, an acute inhalation toxicitystudy in rats demonstrated that administration of Microcyn 60 by thisroute is safe.

The potential irritant effects of Microcyn 60 were evaluated in aprimary ocular irritation study in rabbits. A volume of 0.1 mL ofMicrocyn 60 was instilled in the right eye of three New Zealand whiterabbits. The left eye of each animal was left untreated as a control.The eyes were observed and scored at 1, 24, 48 and 72 hours for cornealulceration or opacity, inflammation of the iris, and redness or chemosisof the conjunctiva. All animals were also observed once daily formortality and signs of ill health.

No signs of ocular irritation were observed in any of the treated orcontrol eyes at any time during the study. All animals appearedclinically healthy for the duration of the study. These findingsindicate that Microcyn 60 does not cause a positive irritation response.

An acute inhalation toxicity study was also performed in rats todetermine the potential inhalation toxicity of Microcyn 60. TenSprauge-Dawley albino rats were exposed to an aerosol generated fromundiluted Microcyn 60 for 4 hours. The concentration of the Microcyn 60was determined to be 2.16 mg/L. All animals were observed frequently onthe day of exposure and once daily for 14 days thereafter for mortalityand clinical/behavioral signs of toxicity. All animals were euthanizedon Day 14 and gross necropsies were performed.

All animals showed very slight to slight piloerection and very slightdecreased activity at 4½ and 6 hours after exposure began but wereasymptomatic by the following day and appeared clinically normal for theduration of the study. One male failed to gain weight between Day 0 andDay 7. There was no mortality and the gross necropsies revealed noobservable abnormalities. The estimated acute inhalation LD50 from thisstudy is greater than 2.16 mg/L.

Additional toxicological studies were performed in the rabbit. Aerosolsuperoxidized water (1 mL) will be delivered to the right nostril via apositive-pressure device to 20 New Zeland rabbits, three times a day for15, 30, 45 and 60 days. The left-control nostril will be left withoutany treatment. Nasomucosa biopsies from the non treated- and M60treated-nostrils will be obtained from five animals at each time point.These tissues will then be observed under optical and electronmicroscopy. A complete medical exam will be conducted in each animalevery other day to document nasal obstruction, facial pain, pressure,mucopurulent rhinorrhea, and malaise. Side effects will be reported asinfrequent, mild, and transient.

Changes to the nasal mucosa appeared after applying intranasal M60 for60 days. There was mild destruction of the epithelia, discreteinflammatory infiltration of the subepithelia region and hyperplasia ofglands and blood vessels in all samples on day 60. Under ultrastructralobservation, we found that varying cyst-like changes within epithelialcells appeared; the mitochondria were condensed and deformed and part ofthe membrane was dissolved. Some epithelial cells were detached;epithelial cilia almost disappeared, and its membrane was dissolved andintercellular spaces were widened. Some cells had detached from thebasement membrane. The tunica propria was mildly edematous.

This study demonstrates that M60 can mildly irritate the nasal mucosaafter intranasal administration for sixty days. However, this damage wasminimum and reversible, so the intranasal route of M60 administrationcould be considered safe. This is based on the fact that although thenasal mucosa can be seriously injured after applying vasoconstrictorsfor several years, it is still restored to normal after stopping thesedrugs. This is possible due to the process of regeneration in the nasalmucosa that depends on whether the basal cells and basement membraneremain intact after injury. Neighboring basal cells can move to thelesion along the basement membrane and cover the lesion. Therefore, evenin the presence of mild detachment of the epithelial cells in someregions after M60 treatment, the basement membrane survived, and thesurviving epithelial cells near the pathological region grew toward theregion lacking the epithelia. Furthermore, topical steroids could havealso been applied to promote recovery of the structure and function ofthe nasal mucosa.

In conclusion, M60 intranasal administration for five days was safe inthis cohort. Pathological mucosa changes were mild and reversible.Therefore, the intranasal administration of M60 could be widely used.

EXAMPLE 12

This example illustrates the activity, stability, and lack of toxicityof an exemplary ORP water.

One such ORP water solution for use in this study is known as“Microcyn,” recently introduced on the Mexican market as an antiseptic.Microcyn is a superoxidized solution of neutral pH with germicidal,sterilizing and wound antiseptic activity in accordance withcertifications obtained from the Secretariat of Health of Mexico.Microcyn is prepared from pure water and salt (NaCl), has a smallconcentration of sodium (<55 ppm) and chlorine (<80 ppm), a pH in therange of 7.2 to 7.8, and oxidation-reduction potential in the range of840 mV to 960 mV. Microcyn is produced in one concentration only, andneed not be activated or diluted.

This solution is produced from water obtained by reverse osmosis, whichis then subjected to an electrochemical gradient generated by highvoltage and sodium chloride. In this way, the reactive species that formin the multiple chambers where the electrochemical gradient is generatedare selected in a controlled way to create Microcyn. The result is asolution with a controlled content of free radicals that confer a highoxidation-reduction potential (+840 mV to +960 mV) and consequently highantimicrobial activity.

Hypochlorous acid and sodium hypochlorite are the most abundant elementscontained in Microcyn, with others in minor concentration, such ashydrogen peroxide, ozone, chloride ions, hydride and sodium hydroxide,among others. Although applicants do not wish to be bound by aparticular theory, it is believed that the disinfectant effect does notnecessarily depend on the quantity of chlorine, but rather, in thecontent of free radicals, since the levels of sodium and chlorine inMicrocyn are less than 50 and 60 parts per million, respectively. Also,and in contrast to other superoxidized solutions that have been reportedin the literature, Microcyn has a neutral pH (6.4-7.8), is not corrosiveand is stable in storage up to 2 years. All these characteristics havemade it possible to produce a superoxidized solution that is effectiveas a high-level disinfectant and compatible for use both on inanimatesurfaces and in tissues.

Accelerated stability tests have demonstrated that Microcyn can bestored in widely varying temperature conditions, from 4 to 65° C.,without losing its disinfectant activity for a period of 2 years. Thisproperty of prolonged stability on the shelf is also the difference fromsuperoxidized solutions reported previously that are only effective ifthey are used immediately after being produced. In other words, whileMicrocyn can be stored and distributed even in extreme conditionswithout losing its antimicrobial activity, other solutions would have tobe produced by a specialized and costly machine in every hospital thattried to use that solution. Nevertheless, the manufacturer recommendsthat, once the container of Microcyn is opened, it be used within 30days for the purpose of guaranteeing uniform activity and consistentresults.

Because Microcyn is produced in only one concentration, the dose ofMicrocyn can be changed only by changes in the volume applied per unitarea of the skin. In the toxicological studies, the doses of Microcynapplied topically to the intact skin varied between 0.05 and 0.07mL/cm²; in the study of acute dermatological toxicity and in theinvestigation of skin irritation, they were up to 8.0 mL/cm², and inthose that investigated its application in deep wounds, Microcyn wasapplied in a dose of 0.09 mL/cm².

Toxicological studies were carried out that applied Microcyn topicallyto the intact skin, using a single application with exposure of 4 to 24h. Multiple applications of Microcyn, one or two times a day, during aperiod of 7 days were assessed for deep wounds in rats.

Two studies were carried out on the intact skin of rabbits to evaluatethe effect of Microcyn as to acute irritation and dermal toxicity. Noclinical signs, dermal irritation, or abnormalities in the skin atautopsy were found in any of the animals exposed to Microcyn.

The characterization of local and systemic toxicity from topicallyapplied Microcyn to a deep wound was evaluated in rats. Noabnormalities, significant differences in the parameters of the bloodchemistry or hematic cytology were observed, nor anomalies in theautopsies. The skin irritation gradings and the histopathology of thewounds and the tissues around the place of application did not revealany difference between the wounds treated with Microcyn and those of thecontrol group treated with saline solution.

The systemic toxicity of Microcyn was also evaluated by means of anintraperitoneal injection in mice. For this, five mice were injectedwith a single dose (50 mL/kg) of Microcyn by the intraperitoneal route.In the same way, five control mice were injected with a single dose (50mL/kg) of saline solution (sodium chloride at 0.9%). In thisinvestigation, neither mortality nor any evidence of systemic toxicitywas observed in any of the animals that received the singleintraperitoneal dose of Microcyn, for which the LD₅₀ is above 50 mL/kg.

Microcyn was administered by the oral route to rats to allow itsabsorption and to characterize any inherent toxic effect of the product.For this a single dose (4.98 mL/kg) was administered by esophageal tubeto three albino rats of the Sprague-Dawley strain. There was nomortality, nor were there clinical signs or abnormalities in theautopsies of any of the animals exposed to the single oral dose ofMicrocyn.

The potential of topically applied Microcyn for ocular irritation wasalso evaluated in rabbits. Ocular irritation was not observed nor anyother clinical sign in any animal exposed to Microcyn by topicaladministration through the ocular route.

Microcyn was applied by the inhalatory route to rats to determinepotential acute toxicity by inhalation. All the animals showed a veryslight or slight reduction in activity and piloerection after theexposure, but they were all asymptomatic on the following day. Mortalityor abnormalities were not observed at autopsy of the animals exposed toMicrocyn by inhalation.

Evaluation of the potential for sensitization of the skin with Microcynwas carried out in guinea pigs using a modified occlusion patch method(Buehler). Irritation was not observed in the animals of the controlgroup after a simple treatment challenge, nor in the animals evaluated(treated by induction) after challenge with the treatment. Therefore,Microcyn does not provoke a sensitizing reaction.

Thus, when it has been applied to the intact skin, deep open dermalwounds, in the conjunctival sac, by oral and inhalation routes or bymeans of intraperitoneal injection, Microcyn has not shown adverseeffects related to the product. There is also experience in havingtreated more than 500 patients with wounds of very diverse nature in theskin and mucosae, with excellent antiseptic and cosmetic results.Accordingly, topically applied Microcyn should be effective andwell-tolerated in this clinical trial.

Microcyn is packaged in transparent 240 mL PET bottles. This product isstored at ambient temperature and remains stable for up to 2 years onthe shelf if the bottle is not opened. On having been opened, it isrecommended that all of the product be used in less than 90 days. Fromits profile of high biological safety, Microcyn can be emptied into thesink without risk of contamination or corrosion.

Multiple microbial trials have been run with Microcyn, both in theUnited States and in Mexico. Eradication of more than 90% of thebacteria occurs in the first few seconds of exposure. The antibacterialand antimycotic activity that Microcyn exhibits in accordance with thisstandard is summarized in Table 5.

TABLE 5 Microcyn Antibacterial and Antimycotic Activity Time of actionBacterium Catalog (reduction below 99.999%) Ps. aeruginosa ATCC 25619 1min St. aureus ATCC 6538 1 min E. coli ATCC 11229 1 min S. typhi CDC 991 min C. albicans ATCC 1 min B. subtilis 9372 Low spore (10⁴) 10 min High spore (10⁶) 15 min 

The sporicidal activity trial was carried out in accordance with thePAHO [Pan-American Health Organization]/WHO protocol.

As for the virucidal activity, Microcyn was found to reduce the viralload of human immunodeficiency virus (strain SF33) by more than 3 logsin five minutes. This was verified by the absence of cytopathic effectand of the antigen Agp24 in the trials of virus treated with Microcyn.These trials were undertaken in accordance with the virucide protocolsof the United States Environmental Protection Agency (DIS/TSS-7/Nov. 12,1981).

The virucidal activity of Microcyn has recently been confirmed instudies carried out in the United States against HIV and polio virus,and its activity against Listeria monocytogenes, MRSA and Mycobacteriumtuberculosis has also been documented. Thus, it has been demonstratedthat Microcyn, when it is administered as recommended, can eradicatebacteria, fungi, viruses and spores from one to fifteen minutes ofexposure.

EXAMPLE 13

This example demonstrates the use of an exemplary ORP water solution,Microcyn as an effective antimicrobial solution.

An In Vitro Time-Kill evaluation was performed using Microcyn oxidativereductive potential water. Microcyn was evaluated versus challengesuspensions of fifty different microorganism strains—twenty-fiveAmerican Type Culture Collection (ATCC) strains and twenty-five ClinicalIsolates of those same species—as described in the Tentative FinalMonograph, Federal Register, 17 Jun. 1994, vol. 59:116, pg. 31444. Thepercent reductions and the Log10 reductions from the initial populationof each challenge strain were determined following exposures to Microcynfor thirty (30) seconds, one (1) minute, three (3) minutes, five (5)minutes, seven (7) minutes, nine (9) minutes, eleven (11) minutes,thirteen (13) minutes, fifteen (15) minutes, and twenty (20) minutes.All agar-plating was performed in duplicate and Microcyn was evaluatedat a 99% (v/v) concentration. All testing was performed in accordancewith Good Laboratory Practices, as specified in 21 C.F.R. Part 58.

The following table summarizes the results of the abovementioned InVitro Time-Kill evaluation at the thirty second exposure mark for allpopulations tested which were reduced by more than 5.0 Log₁₀:

TABLE 6 In Vitro 30-second Kill Initial Post-Exposure PopulationPopulation Log₁₀ Percent No. Microorganism Species (CFU/mL) (CFU/mL)Reduction Reduction 1 Acinetobacter baumannii  2.340 × 10⁹ <1.00 × 10³6.3692 99.9999 (ATCC #19003) 2 Acinetobacter baumannii 1.8150 × 10⁹<1.00 × 10³ 6.2589 99.9999 Clinical Isolate BSLI #061901Ab3 3Bacteroides fragilis  4.40 × 10¹⁰ <1.00 × 10³ 7.6435 99.9999 (ATCC#43858) 4 Bacteroides fragilis  2.70 × 10¹⁰ <1.00 × 10³ 7.4314 99.9999Clinical Isolate BSLI #061901Bf6 5 Candida albicans  2.70 × 10¹⁰ <1.00 ×10³ 6.3345 99.9999 (ATCC #10231) 6 Candida albicans  5.650 × 10⁹ <1.00 ×10³ 6.7520 99.9999 Clinical Isolate BSLI #042905Ca 7 Enterobacteraerogenes 1.2250 × 10⁹ <1.00 × 10³ 6.0881 99.9999 (ATCC #29007) 8Enterobacter aerogenes 1.0150 × 10⁹ <1.00 × 10³ 6.0065 99.9999 ClinicalIsolate BSLI #042905Ea 9 Enterococcus faecalis  2.610 × 10⁹ <1.00 × 10³6.4166 99.9999 (ATCC #29212) 10 Enterococcus faecalis 1.2850 × 10⁹ <1.00× 10³ 6.1089 99.9999 Clinical Isolate BSLI #061901Efs2 11 Enterococcusfaecium  3.250 × 10⁹ <1.00 × 10³ 6.5119 99.9999 VRE, MDR (ATCC #51559)12 Enterococcus faecium  1.130 × 10⁹ <1.00 × 10³ 6.0531 99.9999 ClinicalIsolate BSLI #061901Efm1 13 Escherichia coli  5.00 × 10⁸ <1.00 × 10³5.6990 99.9998 (ATCC #11229) 14 Escherichia coli  3.950 × 10⁸ <1.00 ×10³ 5.5966 99.9997 Clinical Isolate BSLI #042905Ec1 15 Escherichia coli 6.650 × 10⁸ <1.00 × 10³ 5.8228 99.9998 (ATCC #25922) 16 Escherichiacoli  7.40 × 10⁸ <1.00 × 10³ 5.8692 99.9998 Clinical Isolate BSLI#042905Ec2 17 Haemophilus influenzae 1.5050 × 10⁹ <1.00 × 10⁴ 5.177599.9993 (ATCC #8149) 18 Haemophilus influenzae  1.90 × 10⁹ <1.00 × 10⁴5.2788 99.9995 Clinical Isolate BSLI #072605Hi 19 Klebsiella oxytoca 1.120 × 10⁹ <1.00 × 10³ 6.0492 99.9999 MDR (ATCC #15764) 20 Klebsiellaoxytoca  1.810 × 10⁹ <1.00 × 10³ 6.2577 99.9999 Clinical Isolate BSLI#061901Ko1 21 Klebsiella pneumoniae  1.390 × 10⁹ <1.00 × 10³ 6.143099.9999 subsp. ozaenae (ATCC #29019) 22 Klebsiella pneumoniae  9.950 ×10⁸ <1.00 × 10³ 5.9978 99.9999 Clinical Isolate BSLI #061901Kpn2 23Micrococcus luteus  6.950 × 10⁸ <1.00 × 10³ 5.8420 99.9999 (ATCC #7468)24 Micrococcus luteus 1.5150 × 10⁹ <1.00 × 10³ 6.1804 99.9999 ClinicalIsolate BSLI #061901M12 25 Proteus mirabilis 1.5950 × 10⁹ <1.00 × 10³6.2028 99.9999 (ATCC #7002) 26 Proteus mirabilis 2.0950 × 10⁹ <1.00 ×10³ 6.3212 99.9999 Clinical Isolate BSLI #061901Pm2 27 Pseudomonasaeruginosa  6.450 × 10⁸ <1.00 × 10³ 5.8096 99.9999 (ATCC #15442) 28Pseudomonas aeruginosa 1.3850 × 10⁹ <1.00 × 10³ 6.1414 99.9999 ClinicalIsolate BSLI #072605Pa 29 Pseudomonas aeruginosa  5.550 × 10⁸ <1.00 ×10³ 5.7443 99.9999 (ATCC #27853) 30 Pseudomonas aeruginosa 1.1650 × 10⁹<1.00 × 10³ 6.0663 99.9999 Clinical Isolate BSLI #061901Pa2 31 Serratiamarcescens  9.950 × 10⁸ <1.00 × 10³ 5.9978 99.9999 (ATCC #14756) 32Serratia marcescens 3.6650 × 10⁹ <1.00 × 10³ 6.5641 99.9999 ClinicalIsolate BSLI #042905Sm 33 Staphylococcus aureus 1.5050 × 10⁹ <1.00 × 10³6.1775 99.9999 (ATCC #6538) 34 Staphylococcus aureus  1.250 × 10⁹ <1.00× 10³ 6.0969 99.9999 Clinical Isolate BSLI #061901Sa1 35 Staphylococcusaureus  1.740 × 10⁹ <1.00 × 10³ 6.2405 99.9999 (ATCC #29213) 36Staphylococcus aureus 1.1050 × 10⁹ <1.00 × 10³ 6.0434 99.9999 ClinicalIsolate BSLI #061901Sa2 37 Staphylococcus epidermidis 1.0550 × 10⁹ <1.00× 10³ 6.0233 99.9999 (ATCC #12228) 38 Staphylococcus epidermidis  4.350× 10⁸ <1.00 × 10³ 5.6385 99.9998 Clinical Isolate BSLI #072605Se 39Staphylococcus haemolyticus  8.150 × 10⁸ <1.00 × 10³ 5.9112 99.9999(ATCC #29970) 40 Staphylococcus haemolyticus  8.350 × 10⁸ <1.00 × 10³5.9217 99.9999 Clinical Isolate BSLI #042905Sha 41 Staphylococcushominis  2.790 × 10⁸ <1.00 × 10³ 5.4456 99.9996 (ATCC #27844) 42Staphylococcus hominis  5.20 × 10⁸ <1.00 × 10³ 5.7160 99.9998 ClinicalIsolate BSLI #042905Sho 43 Staphylococcus saprophyticus  9.10 × 10⁸<1.00 × 10³ 5.9590 99.9999 (ATCC #35552) 44 Staphylococcus saprophyticus1.4150 × 10⁹ <1.00 × 10³ 6.1508 99.9999 Clinical Isolate BSLI #042905Ss45 Streptococcus pneumoniae 2.1450 × 10⁹ <1.00 × 10⁴ 5.3314 99.9995(ATCC #33400) 46 Streptococcus pyogenes  5.20 × 10⁹ <1.00 × 10³ 6.716099.9999 (ATCC #19615) 47 Streptococcus pyogenes 2.5920 × 10⁹ <1.00 × 10³6.4141 99.9999 Clinical Isolate BSLI #061901Spy7

While their microbial reductions were measured at less than 5.0 Log₁₀,Microcyn also demonstrated antimicrobial activity against the remainingthree species not included in Table 6. More specifically, a thirtysecond exposure to Microcyn reduced the population of Streptococcuspneumoniae (Clinical Isolate; BSLI #072605Spn1) by more than 4.5 Log₁₀,which was the limit of detection versus this species. Further, whenchallenged with Candida tropicalis (ATCC #750), Microcyn demonstrated amicrobial reduction in excess of 3.0 Log₁₀ following a thirty secondexposure. Additionally, when challenged with Candida tropicalis (BSLI#042905Ct), Microcyn demonstrated a microbial reduction in excess of 3.0Log₁₀ following a twenty minute exposure.

The exemplary results of this In Vitro Time-Kill evaluation demonstratethat Microcyn oxidative reductive potential water exhibits rapid (i.e.,less than 30 seconds in most cases) antimicrobial activity versus abroad spectrum of challenging microorganisms. Microbial populations offorty-seven out of the fifty Gram-positive, Gram-negative, and yeastspecies evaluated were reduced by more than 5.0 Log₁₀ within thirtyseconds of exposure to the product.

EXAMPLE 14

This example demonstrates a comparison of the antimicrobial activity ofan exemplary ORP water solution, Microcyn, versus HIBICLENS®chlorhexidine gluconate solution 4.0% (w/v) and 0.9% sodium chlorideirrigation (USP).

An In Vitro Time-Kill evaluation was performed as described in Example13 using HIBICLENS® chlorhexidine gluconate solution 4.0% (w/v) and asterile 0.9% sodium chloride irrigation solution (USP) as referenceproducts. Each reference product was evaluated versus suspensions of theten American Type Culture Collection (ATCC) strains specifically denotedin the Tentative Final Monograph. The data collected was then analyzedagainst the Microcyn microbial reduction activity recorded in Example13.

Microcyn oxidative reductive potential water reduced microbialpopulations of five of the challenge strains to a level comparable tothat observed for the HIBICLENS® chlorhexidine gluconate solution. BothMicrocyn and HIBICLENS® provided a microbial reduction of more than 5.0Log₁₀ following a thirty second exposure to the following species:Escherichia coli (ATCC #11229 and ATCC #25922), Pseudomonas aeruginosa(ATCC #15442 and ATCC #27853), and Serratia marcescens (ATCC #14756).Further, as shown above in Table 5, Microcyn demonstrated excellentantimicrobial activity against Micrococcus luteus (ATCC #7468) byproviding a 5.8420 Log₁₀ reduction after a thirty second exposure.However, a direct Micrococcus luteus (ATCC #7468) activity comparison toHIBICLENS® was not possible because after a thirty second exposure,HIBICLENS® reduced the population by the detection limit of the test (inthis specific case, by more than 4.8 Log₁₀). It is noted that thesterile 0.9% sodium chloride irrigation solution reduced microbialpopulations of each of the six challenge strains discussed above by lessthan 0.3 Log₁₀ following a full twenty minute exposure.

Microcyn oxidative reductive potential water provided greaterantimicrobial activity than both HIBICLENS® and the sodium chlorideirrigation for four of the challenge strains tested: Enterococcusfaecalis (ATCC #29212), Staphylococcus aureus (ATCC #6538 and ATCC#29213), and Staphylococcus epidermidis (ATCC #12228). The followingtable summarizes the microbial reduction results of the In VitroTime-Kill evaluation for these four species:

TABLE 7 Comparative Kill Results Log₁₀ Reduction Microorganism ExposureNaCl Species Time Microcyn HIBICLENS ® Irrigation Enterococcus 30seconds 6.4166 1.6004 0.3180 faecalis 1 minute 6.4166 2.4648 0.2478(ATCC #29212) 3 minutes 6.4166 5.2405 0.2376 5 minutes 6.4166 5.41660.2305 7 minutes 6.4166 5.4166 0.2736 9 minutes 6.4166 5.4166 0.2895 11minutes 6.4166 5.4166 0.2221 13 minutes 6.4166 5.4166 0.2783 15 minutes6.4166 5.4166 0.2098 20 minutes 6.4166 5.4166 0.2847 Staphylococcus 30seconds 6.1775 1.1130 0.0000 aureus 1 minute 6.1775 1.7650 0.0191 (ATCC#6538) 3 minutes 6.1775 4.3024 0.0000 5 minutes 6.1775 5.1775 0.0000 7minutes 6.1775 5.1775 0.0000 9 minutes 6.1775 5.1775 0.0000 11 minutes6.1775 5.1775 0.0267 13 minutes 6.1775 5.1775 0.0000 15 minutes 6.17755.1775 0.0191 20 minutes 6.1775 5.1775 0.0000 Staphylococcus 30 seconds6.2405 0.9309 0.0000 aureus 1 minute 6.2405 1.6173 0.0000 (ATCC #29213)3 minutes 6.2405 3.8091 0.0460 5 minutes 6.2405 5.2405 0.0139 7 minutes6.2405 5.2405 0.0000 9 minutes 6.2405 5.2405 0.0113 11 minutes 6.24055.2405 0.0283 13 minutes 6.2405 5.2405 0.0000 15 minutes 6.2405 5.24050.0000 20 minutes 6.2405 5.2405 0.0615 Staphylococcus 30 seconds 5.63855.0233 0.0456 epidermidis 1 minute 5.6385 5.0233 0.0410 (ATCC #12228) 3minutes 5.6385 5.0233 0.0715 5 minutes 5.6385 5.0233 0.0888 7 minutes5.6385 5.0233 0.0063 9 minutes 5.6385 5.0233 0.0643 11 minutes 5.63855.0233 0.0211 13 minutes 5.6385 5.0233 0.1121 15 minutes 5.6385 5.02330.0321 20 minutes 5.6385 5.0233 0.1042

The results of this comparative In Vitro Time-Kill evaluationdemonstrate that Microcyn oxidative reductive potential water not onlyexhibits comparable antimicrobial activity to HIBICLENS® againstEscherichia coli (ATCC #11229 and ATCC #25922), Pseudomonas aeruginosa(ATCC #15442 and ATCC #27853), Serratia marcescens (ATCC #14756), andMicrococcus luteus (ATCC #7468), but provides more effective treatmentagainst Enterococcus faecalis (ATCC #29212), Staphylococcus aureus (ATCC#6538 and ATCC #29213), and Staphylococcus epidermidis (ATCC #12228). Asshown in Table 7, Microcyn exemplifies a more rapid antimicrobialresponse (i.e., less than 30 seconds) in some species. Moreover,exposure to Microcyn results in a greater overall microbial reduction inall species listed in Table 7.

EXAMPLE 15

This example provides a formulation of the invention suitable fortopical administration to a patient. The formulation contains thefollowing:

Component Quantity ORP water solution 250 mL Carbopol ® polymer powder(thickening agent) 15 g Triethanolamine (neutralizing agent) 80 mL

EXAMPLE 16

This example provides a formulation of the invention suitable fortopical administration to a patient. The formulation contains thefollowing:

Component Quantity ORP water solution 1000 mL Carbopol ® polymer powder(thickening agent) 15 g Triethanolamine (neutralizing agent) 80 mL

EXAMPLE 17

This example provides a formulation of the invention suitable fortopical administration to a patient. The formulation contains thefollowing:

Component Quantity ORP water solution 250 mL Carbopol ® polymer powder(thickening agent) 7 g Triethanolamine (neutralizing agent) 12 mL

EXAMPLE 18

This example describes the manufacture of a formulation of the inventioncomprising an ORP water solution and a thickening agent.

An ORP water solution is put into a suitable container, such as a glassbeaker or jar. Carbopol® 974P polymer is passed through a coarse sieve(or strainer), which permits rapid sprinkling, whilst at the same timebreaking up any large agglomerates. The polymer Carbopol® 974P is thenadded as the thickening agent. The Carbopol®D polymer is added slowly toprevent the formation of clumps and, thus, avoid an excessively longmixing cycle.

The solution is mixed rapidly during the addition of the Carbopol®polymer so that the powder dissolves at room temperature. Theneutralizing agent triethanolamine is then added to the solution andmixed by means of an electric mixer or other suitable device, until ahomogeneous gel is obtained. The addition of the neutralizing agent tothe Carbopol® polymer composition converts the formulation into a gel.

EXAMPLE 19

This example describes the use of ORP water solution according to thepresent invention for the treatment of burns, particularly second, andthird degree burns, in pediatric burn patients.

A total of 64 human pediatric burn patients were treated with an ORPwater solution. The study group was compared to a control group alsoconsisting of 64 patients treated with conventional burn therapy. Thestudy group consisted of the following: 1 patient with first degreeburns, 6 patients with a combination of first and second degree burns,38 patients with second degree burns, 4 patients with third degreeburns, and 15 patients with a combination of second and third degreeburns. Moreover, the study group consisted of patients having burns overthe following percent of their bodies (i.e., extension of the burn): 10patients with 0 to 9% extension of the burn, 27 patients with 10 to 19%extension of the burn, 11 patients with 20 to 29% extension of the burn,8 patients with 30 to 39% extension of the burn, 4 patients with 40 to49% extension of the burn, 1 patients with 50 to 59% extension of theburn, and 3 patients with 60 to 69% extension of the burn. Each burn wasinitially debribed. The solution was applied by spraying with highpressure irrigation device. Next, the solution was applied by sprayingand left to moisten the burn for 5 to 15 minutes, which was repeatedthree times a day. The burns were not dressed in between administrationof the solution.

In cultures taken to determine the presence of microorganisms on thesurface of the burn, only 6 patients treated with the ORP water solutionhad a positive culture after 7-15 days at the hospital, compared to 22patients in the control group. The remaining patients in the study group(58) and control group (42) had negative cultures. The microorganismspresent in the positive cultures from the study and control groups areset forth in Table 8.

TABLE 8 Burn Microbiology Control Group % Study Group % Staph. aureus56.0 Staph. aureus 57.1 Enterobacter 8.0 Enterobacter 28.2 cloacaecloacae Staph. haemolyticus 14.2 Pseudomonas 19.0 aeruginosa Candidaalbicans 12.0 Klebsiella sp. 5.0 Total 100.0 Total 100.0

The frequency of application of the ORP water solution varied accordingto the nature of each patient's burn. The average hospital stay by burngrade for the study group and control group was tabulated. For firstdegree burns, the average hospital stay was 4.6 days for the study group(6 patients) compared to 19.2 days for the control group (45 patients).For second degree burns, the average hospital stay was 10.6 days for thestudy group (44 patients) as compared to 26.9 days for the control group(9 patients). For third degree burns, the average hospital stay was 29.5days for the study group (14 patients) as compared to 39.8 days for thecontrol group (10 patients). Overall, the length of the average hospitalstay was reduced by 48% from 28.6 days to 14.9 days with theadministration of the ORP water solution of the invention to pediatricburn patients. The average stay in the hospital in number of days forthe control group v. the study group based on the extent of the burn isset forth in Table 9.

TABLE 9 Hospital Stay Hospital Stay in Days Hospital Stay in DaysExtension of Burn Control Group Study Group 0 to 9% 16.1 6.9 10 to 19%11.7 8.2 20 to 29%  8.6 22.7 30 to 39% 40.2 16.8 40 to 49% 32.3 26.5 50to 59% 0 (no patients treated) 55 60 to 69% 34.3 68.0

As is apparent from this example, the ORP water solution of the presentinvention can advantageously be administered to pediatric burn patientsresulting in reduced hospital stays.

EXAMPLE 20

This example describes the administration of the ORP water solution ofthe present invention to pediatric burn patients without theadministration of antibiotics.

None of the 58 patients in the study group who had negativemicroorganism cultures measured after 7-15 days in the hospitaldescribed in Example 19 above were treated with antibiotics. The averagehospital stay for this group of patients was 12.3 days. In the controlgroup, antibiotics were used on 46 patients in addition to theadministration of the ORP water solution. Positive cultures formicroorganisms were observed in 22 of these patients with an averagehospital stay of 28.6 days for the patients using antibiotics.

As is apparent from this example, the ORP water solution of the presentinvention can advantageously be administered to pediatric burn patientswithout the routine use of antibiotics.

EXAMPLE 21

This example demonstrates the effect of an exemplary ORP water solutionversus hydrogen peroxide (HP) on the viability of human diploidfibroblasts (HDFs). To study this potential toxicity, HDFs were exposedin vitro to ORP water solution and hydrogen peroxide (HP). HP is knownto be toxic to eukaryotic cells, increasing apoptosis and necrosis andreducing cellular viability. In this example, cell viability, apoptosisand necrosis were measured in HDFs exposed to pure ORP water solutionand 880 mM HP (a concentration employed for antiseptic uses of HP) for 5and 30 minutes.

HDF cultures were obtained from three different foreskins, which werepooled and cryopreserved together for the purpose of this study. Onlydiploid cells were used for all experiments. On cell cycle analysis, DNAdiploidy was defined as the presence of a single G0-G1 peak with a CV≦7%and a corresponding G2/M peak collected from at least 20,000 totalevents. FIGS. 4A-4C discloses the results with exposure times of 5 and30 minutes are depicted in white and black bars, respectively.Simultaneous analyses of these parameters were performed in the samecell populations by flow cytometry using: A) 7-aminoactinomycin D(7AAD); B) Annexin V-FITC and C) Propidium iodide. FIGS. 8A-8C disclosepercentage values expressed as mean±SD (n=3).

Cell viability was 75% and 55% after a 5 minute exposure to ORP watersolution and HP, respectively (FIG. 4A). If the exposure was prolongedto 30 min, cell viability further decreased to 60% and 5%, respectively.Apparently, the ORP water solution induced cell death through necrosisbecause 15% of the cells incorporated propidium iodide in the flowcytometry analysis at both times (FIG. 4C). While not wanting to bebound by any particular theory, this result could be due to an osmoticeffect induced by the hypotonicity of Microcyn (13mOsm) since the cellswere kept in the ORP water solution only, without added growth factorsor ions. Apoptosis does not seem to be the mechanism by which the ORPwater solution induces cell death because only 3% of ORP watersolution-treated cells exposed Annexin-V in the cellular surface (amarker of apoptosis) (FIG. 4B). This percentage was actually similar tothe one measured in the control group. On the contrary, HP inducednecrosis in 20% and 75% of treated cells and apoptosis in 15% and 20%after 5 and 30 min of exposure, respectively. Altogether these resultsshow that the (undiluted) ORP water solution is far less toxic for HDFsthan an antiseptic concentration of HP.

EXAMPLE 22

This example demonstrates the effect of an exemplary ORP water solutionrelative to hydrogen peroxide (HP) on oxidative DNA damage and formationof the DNA adduct 8-hydroxy-2′-deoxiguanosine (8-OHdG) in HDFs. It isknown that the production of 8-OHdG adducts in a cell is a marker ofoxidative damage at specific residues of DNA. In addition, high cellularlevels of this adduct correlate with mutagenesis, carcinogenesis andcellular aging.

FIG. 5 shows the levels of 8-OHdG adducts present in DNA samples fromHDFs after control treatments, ORP water solution treatments andHP-treatments for 30 minutes. DNA was extracted right after the exposure(TO, white bars) or three hours after the challenge period (T3, blackbars). DNA was digested and the 8-OHdG adducts were measured by ELISAkit as per the manufacturer's instructions. Values are shown (ng/mL) asmean±SD (n=3). The exposure to ORP water solution for 30 minutes did notincrease the formation of adducts in the treated cells in comparison tocontrol cells after incubation for 30 minutes. In contrast, thetreatment with highly diluted HP—down to sublethal and nontherapeutic HPconcentrations (500 μM HP)—the treatment with 500 μM HP for 30 minutesincreased the number of 8-OHdG adducts by about 25 fold relative to thecontrol-treated or ORP water solution-treated cells.

The ORP water solution-treated cells were able to decrease the levels of8-OHdG adducts if left in supplemented DMEM for 3 hours after exposureto the ORP water solution. Despite being allowed the same 3 hourrecovery period, HP-treated cells still presented about 5 times moreadducts than control-treated or ORP water solution treated cells.Altogether, these results demonstrate that acute exposure to the ORPwater solution does not induce significant DNA oxidative damage. Theseresults also indicate that the ORP water solution will not likely inducemutagenesis or carcinogenesis in vitro or in vivo.

EXAMPLE 23

This example demonstrates the effects on HDFs of chronic exposure to lowconcentrations of an exemplary ORP water solution versus HP. It is knownthat chronic oxidative stress induces premature aging of cells. In orderto mimic a prolonged oxidative stress, primary HDF cultures werechronically exposed to a low concentration of the ORP water solution(10%) or a non lethal-HP concentration (5 μM) during 20 populationdoublings. The expression and activity of the SA-β-galactosidase enzymehas previously been associated with the senescence process in vivo andin vitro. In this example the expression of the SA-β-galactosidaseenzyme was analyzed after one month of continuous exposure of HDF to theORP water solution or HP. The results are depicted in FIG. 6. Theexpression of the enzyme SA-β-galactosidase was analyzed by counting thenumber of blue cells in 20 microscopic fields. (For an example stainingpattern, see Panel A.) Panel B shows that only HP treatment acceleratedthe aging of cells as indicated by the number of cells over-expressingSA-β-galactosidase (n=3). Chronic treatment with a low dose of HPincreased the SA-β-Gal expression in 86% of cells while the treatmentwith the ORP water solution did not induce the overexpression of thisprotein. It can be concluded from this example that ORP water solutionis not an inducer of premature cellular aging.

EXAMPLE 24

This example demonstrates the results of a toxicity study using anexemplary ORP water solution.

An acute systemic toxicity study was performed in mice to determine thepotential systemic toxicity of Microcyn 60, an exemplary ORP watersolution. A single dose (50 mL/kg) of Microcyn 60 was injectedintraperitoneally in five mice. Five control mice were injected with asingle dose (50 mL/kg) of saline (0.9% sodium chloride). All animalswere observed for mortality and adverse reactions immediately followingthe injection, at 4 hours after injection, and then once daily for 7days. All animals were also weighed prior to the injection and again onDay 7. There was no mortality during the study. All animals appearedclinically normal throughout the study. All animals gained weight. Theestimated Microcyn 60 acute intraperitoneal LD50 from this study isgreater than 50 mL/kg. This example demonstrates that Microcyn 60 lackssignificant toxicity and should be safe for therapeutic use accordancewith the invention.

EXAMPLE 25

This example illustrates a study conducted to determine the potentialcytogenetic toxicity of an exemplary ORP water solution.

A micronucleus test was performed using an exemplary ORP water solution(Microcyn 10%) to evaluate the mutagenic potential of intraperitonealinjection of an ORP water solution into mice. The mammalian in vivomicronucleus test is used for the identification of substances whichcause damage to chromosomes or the mitotic apparatus of murinepolychromatic erythrocytes. This damage results in the formation of“micronuclei,” intracellular structures containing lagging chromosomefragments or isolated whole chromosomes. The ORP water solution studyincluded 3 groups of 10 mice each (5 males/5 females): a test group,dosed with the ORP water solution; a negative control group, dosed witha 0.9% NaCl solution; and a positive control group, dosed with amutagenic cyclophosphamide solution. The test and the negative controlgroups received an intraperitoneal injection (12.5 ml/kg) of the ORPwater solution or 0.9% NaCl solution, respectively, for two consecutivedays (days 1 and 2). The positive control mice received a singleintraperitoneal injection of cyclophosphamide (8 mg/mL, 12.5 ml/kg) onday 2. All mice were observed immediately after injection for anyadverse reactions. All animals appeared clinically normal throughout thestudy and no sign of toxicity was noted in any group. On day 3, all micewere weighed and terminated.

The femurs were excised from the terminated mice, the bone marrow wasextracted, and duplicate smear preparations were performed for eachmouse. The bone marrow slides for each animal were read at 40×magnification. The ratio of polychromatic erythrocytes (PCE) tonormochromatic erythrocytes (NCE), an index of bone marrow toxicity, wasdetermined for each mouse by counting a total of at least 200erythrocytes. Then a minimum of 2000 scoreable PCE per mouse wereevaluated for the incidence of micronucleated polychromaticerythrocytes. Statistical analysis of the data were done using the Mannand Whitney test (at 5% risk threshold) from a statistical softwarepackage (Statview 5.0, SAS Institute Inc., USA).

The positive control mice had statistically significant lower PCE/NCEratios when compared to their respective negative controls (males: 0.77vs. 0.90 and females: 0.73 vs. 1.02), showing the toxicity of thecyclophosphamide on treated bone marrow. However, there was nostatistically significant difference between the PCE/NCE ratios for theORP water solution-treated mice and negative controls. Similarly,positive control mice had a statistically significant higher number ofpolychromatic erythrocytes bearing micronuclei as compared to both theORP water solution-treated mice (males: 11.0 vs. 1.4/females: 12.6 vs.0.8) and the negative controls (males: 11.0 vs. 0.6/females: 12.6 vs.1.0). There was no statistically significant difference between thenumber of polychromatic erythrocytes bearing micronculei in ORP watersolution-treated and negative control mice.

This example demonstrates that Microcyn 10% did not induce toxicity ormutagenic effects after intraperitoneal injections into mice.

EXAMPLE 26

This study demonstrates the lack of toxicity of an exemplary ORP watersolution, Dermacyn.

This study was done in accordance with ISO 10993-5:1999 standard todetermine the potential of an exemplary ORP water solution, Dermacyn, tocause cytotoxicity. A filter disc with 0.1 mL of Dermacyn was placedonto an agarose surface, directly overlaying a monolayer of mousefibroblast cells (L-929). The prepared samples were observed forcytotoxic damage after 24 hours of incubation at 37° C. in the presenceof 5% CO₂. Observations were compared to positive and negative controlsamples. The Dermacyn containing samples did not reveal any evidence ofcell lysis or toxicity, while positive and negative control performed asanticipated.

Based on this study Dermacyn was concluded not to generate cytotoxiceffects on murine fibroblasts.

EXAMPLE 27

This study was conducted with 16 rats to evaluate the local tolerabilityof an exemplary ORP water solution, Dermacyn, and its effects on thehistopathology of wound beds in a model of full-thickness dermal woundhealing. Wounds were made on both sides of the subject rat. During thehealing process skin sections were taken on either the left or the rightsides (e.g., Dermacyn-treated and saline-treated, respectively).

Masson's trichrome-stained sections and Collagen Type II stainedsections of the Dermacyn and saline-treated surgical wound sites wereevaluated by a board-certified veterinary pathologist. The sections wereassessed for the amount of Collogen Type 2 expression as amanifestiation of connective tissue proliferation, fibroblast morphologyand collagen formation, presence of neoepidermis in cross section,inflammation and extent of dermal ulceration.

The findings indicate that Dermacyn was well tolerated in rats. Therewere no treatment-related histopathologic lesions in the skin sectionsfrom either sides' wounds (Dermacyn-treated and saline-treated,respectively). There were no relevant histopathologic differencesbetween the saline-treated and the Dermacyn-treated wound sites,indicating that the Dermacyn-treatement was well tolerated. There wereno significant differences between Collagen Type 2 expression betweenthe saline-treated and the Dermacyn-treated wound sites indicating thatthe Dermacyn does not have an adverse effect on fibroblasts or oncollagen elaboration during wound healing.

EXAMPLE 28

This study can be done to demonstrate the safety and efficacy of anexemplary ORP water solution, Dermacyn, used in accordance with theinvention as a replacement solution for the Versajet™ (Smith & Nephew)jet lavage system in the treatment of necrotic tissue (ulcers) distal tothe malleoli, as compared to the standard regimen.

This will be a prospective randomized, double-blind, controlled study.Approximately 30 patients (about 20 in the Dermacyn group/about 10 inthe Control group) will be enrolled in the study. The population forthis study will be patients with lower extremity ulcers (e.g., diabeticfoot ulcers, venous stasis ulcers). All of the study's inclusion andexclusion criteria must be satisfied by the Day 0 for the patient to beeligible for enrollment into the study. The inclusion criteria are:patient is 18 years old or older; patient's lower extremity ulcer hasnecrotic tissue present and is a candidate for mechanical debridement bythe jet lavage system; patient's ulcer is located distal to themalleoli; patient's ulcer surface area is greater than or equal to 1.0cm²; patient's ulcer extends through the dermis and into subcutaneoustissue (granulation tissue may be present), with possible exposure ofmuscle, or tendon, but without bone, and/or joint capsule involvement;and patient's Ankle-Arm Index by Doppler is an ABI of greater than orequal to 0.8 or patient's toe pressure is greater than or equal to 40mmHg.

The exclusion criteria are: patient has clinical evidence of gangrene onany part of the treatment limb; patient's ulcer is expected to beresected or amputated during the study period; patient's has thefollowing signs of a systemic inflammatory response syndrome (SIRS);patient's ulcer has a total surface area that is less than 1 cm²;patient has one or more medical condition(s) (including renal, hepatic,hematologic, neurologic, or immune disease) that in the opinion of theinvestigator would make the patient an inappropriate candidate for thisstudy; patient has known active alcohol or drug abuse; patient isreceiving oral or parenteral corticosteroids, immunosuppressive orcytotoxic agents, or is anticipated to require such agents during thecourse of the study; patient has known allergies to chlorine; patient'sulcer is accompanied by osteomyelitis; and patient has any condition(s)which seriously compromises the patient's ability to complete thisstudy.

After the informed consent has been obtained, inclusion and exclusioncriteria met, the patient will be randomized (2:1 randomization) intoone of the following treatments: Treatment—Dermacyn with the jet lavagesystem, plus the use of a hydrogel wound dressing regimen;Control—Saline (standard treatment with the jet lavage systems), plusthe use of a hydrogel wound dressing regimen.

Each patient randomized to Dermacyn will receive applications of thestudy product Dermacyn, with the Versajet jet lavage system duringmechanical debridement of the patient's wound. A standard pressuresetting on the Versajet will be used for diabetic foot ulcers, whichwill be distal to the malleoli. After debridement, Dermacyn will beapplied onto the wound in sufficient quantities to rinse the wound bedfree of debris. The wound will be covered with a hydrogel dressing. Atevery dressing change, the wound will be rinsed out with Dermacyn andcovered with a new hydrogel dressing. The dressings will be changedevery 3 days, unless otherwise specified by the investigator. Theclinical response factors (CFRs) ((1) reduction of bacteria in thewound, (2) reduction in wound area, and (3) development of granulationtissue) will be determined during the weekly visits.

Each Control patient will receive applications of the Control product(saline solution) with the Versajet jet lavage system during mechanicaldebridement of the patient's wound. After debridement, saline will beapplied onto the wound in sufficient quantities to rinse the wound bedfree of debris. The wound will be covered with a hydrogel dressing. Atevery dressing change, the wound will be rinsed out with saline andcovered with a new hydrogel dressing. The dressings will be changedevery 3 days, unless otherwise specified by the investigator. Theclinical response factors will be determined during the weekly visits.

Debridement of the wound may be performed at each weekly visit. Anynecrotic tissue will be debrided with jet lavage prior to the woundassessments. Debris from the ulcer will be rinsed with either Dermacynor saline (dependent upon the randomization). Between visits the patientwill rinse the wound with Dermacyn or saline (dependent uponrandomization) at every dressing change. Photographs of the wound willbe taken at every visit after debridement.

The primary efficacy endpoints will be: (1) reduction of bacteria in thewound, (2) reduction in wound area, and (3) development of granulationtissue. Safety will be assessed in all patients who are randomized inthe study. The treatment of emergent and serious adverse events will berecorded.

EXAMPLE 29

This study will demonstrate the safety and efficacy of an exemplary ORPwater solution, Dermacyn, as a replacement solution for the Jet-Ox NDlavage system in the treatment of necrotic tissue in lower extremityulcers as compared to the standard regimen used by the Jet-Ox ND system.

The Jet-Ox ND system removes necrotic tissue from chronic wounds via acontrolled spray lavage of sterile saline, without damage to underlyinghealthy tissues. This study will replace saline with Dermacyn, which isexpected to provide the same spray lavage effect and additionally reducethe bacterial load of the wound that may be inhibiting wound closure.

Twenty patients will be studied (randomized to yield 10 Dermacynpatients and 10 Control patients). The inclusion criteria will be:patient is older than 18 years; patient has a lower extremitybelow-the-knee ulcer with necrotic tissue present and is a candidate formechanical debridement with the Jet-Ox ND lavage system; patients ulcerhas been present >30 days prior to the screening visit; the ulcersurface area is >1 cm2 the ulcer extends through the dermis and intosubcutaneous tissue (granulation tissue may be present) with possibleexposure of muscle, tendon but, without exposed bone or capsule;patients ankle/arm index by doppler is >0.8 and/or patients toe pressureis >40 mmHg; and the patient has a palpable pulse at the dorsalis pedisand/or posterior tibial artery.

There will be the following exclusion criteria: renal, hepatic,hematologic, neurologic or immuno-compromised patients, including havingHuman Immunodeficiency virus (HIV) or Acquired Immunodeficiency Syndrome(AIDS); that in the opinion of the investigator would make the patientan inappropriate candidate for the study; wounds with the followingclinical signs of infection; gangrene on any part of the treatment limb;ulcer exhibits exposed bone (positive probe to bone) or has otherevidence of underlying osteomyelitis at the ulcer site; expectation thatthe infected ulcer will be amputated or resected during the studyperiod; severe malnutrition as evidenced by an albumin of <2.0; knownalcohol or drug abuse; patients receiving oral or parenteralcorticosteroids, immunosuppressive or cytotoxic agents, coumadin,heparin, or is anticipated to require such agents during the course ofthe study; and patient has known allergy to chlorine.

Each individual will be randomized into one of two treatment arms;Dermacyn or saline. The target ulcer will receive mechanicaldebridement, followed by irrigation of the wound with either Dermacyn orsaline and bandaging with a hydrogel dressing. A central wound biopsyfor quantitive culture will be taken, along with laboratory studies(hematology, serum chemistry and pregnancy testing as appropriate),non-invasive peripheral vascular studies, medical history and physicalexamination, ulcer tracings, and ulcer photographs.

A Jet-Ox ND lavage system will be dispensed along with Dermacyn orsaline, hydrogel and bandaging materials. Directions for home use willbe provided. Visits will include screening, enrollment [day 0] withrandomization, weekly visits with debridement, photographs andassessments. Efficacy will be determined by (1) reduction of bacteria inthe wound, (2) reduction in wound area, and (3) development ofgranulation tissue during the course of the study. Safety will beassessed in all patients who are randomized in the study. Treatmentemergent and serious adverse events will be recorded.

All references, including publications, patent applications, andpatents, cited herein are hereby incorporated by reference to the sameextent as if each reference were individually and specifically indicatedto be incorporated by reference and were set forth in its entiretyherein.

The use of the terms “a” and “an” and “the” and similar referents in thecontext of describing the invention (especially in the context of thefollowing claims) are to be construed to cover both the singular and theplural, unless otherwise indicated herein or clearly contradicted bycontext. The terms “comprising,” “having,” “including,” and “containing”are to be construed as open-ended terms (i.e., meaning “including, butnot limited to,”) unless otherwise noted. Recitation of ranges of valuesherein are merely intended to serve as a shorthand method of referringindividually to each separate value falling within the range, unlessotherwise indicated herein, and each separate value is incorporated intothe specification as if it were individually recited herein. All methodsdescribed herein can be performed in any suitable order unless otherwiseindicated herein or otherwise clearly contradicted by context. The useof any and all examples, or exemplary language (e.g., “such as”)provided herein, is intended merely to better illuminate the inventionand does not pose a limitation on the scope of the invention unlessotherwise claimed. No language in the specification should be construedas indicating any non-claimed element as essential to the practice ofthe invention.

Preferred embodiments of this invention are described herein, includingthe best mode known to the inventors for carrying out the invention.Variations of those preferred embodiments may become apparent to thoseof ordinary skill in the art upon reading the foregoing description. Theinventors expect skilled artisans to employ such variations asappropriate, and the inventors intend for the invention to be practicedotherwise than as specifically described herein. Accordingly, thisinvention includes all modifications and equivalents of the subjectmatter recited in the claims appended hereto as permitted by applicablelaw. Moreover, any combination of the above-described elements in allpossible variations thereof is encompassed by the invention unlessotherwise indicated herein or otherwise clearly contradicted by context.

What is claimed is:
 1. A method of treating a second or third degreeburn in a patient, comprising: administering an oxidative reductivepotential water solution to the patient in an amount sufficient to treatthe burn, wherein the solution has a pH of 7.4 to 7.6, wherein thesolution is stable for at least two months, wherein the solutioncomprises anode water and cathode water, wherein the oxidative reductivepotential water solution comprises free chlorine species at a level offrom 50 ppm to 80 ppm, wherein the free chlorine species is selectedfrom the group selected from the group consisting of hypochlorous acid,hypochlorite ions, sodium hypochlorite, chlorite ions, chloride ions,dissolved chlorine gas, and mixtures thereof, and wherein the freechlorine species comprises at least one of: hypochlorous acid in anamount of 15 ppm to 35 ppm and sodium hypochlorite in an amount of 25ppm to 50 ppm.
 2. The method of claim 1, wherein the cathode water ispresent in an amount of from 10% by volume to 50% by volume of thesolution.
 3. The method of claim 2, wherein the cathode water is presentin an amount of from 20% by volume to 40% by volume of the solution. 4.The method of claim 2, wherein the anode water is present in an amountof from 50% by volume to 90% by volume of the solution.
 5. The method ofclaim 1, wherein the solution is administered to the patient by sprayingthe burn with the solution.
 6. The method of claim 5, wherein thesolution is administered to the patient by spraying the burn with thesolution with a high pressure irrigation device.
 7. The method of claim5, wherein the burn is moistened with the solution for at least fiveminutes.
 8. The method of claim 7, wherein the burn is moistened withthe solution for at least 15 minutes.
 9. The method of claim 1, whereinthe solution is administered to the patient at least daily.
 10. Themethod of claim 9, wherein the solution is administered to the patientthree times a day.
 11. A method of treating a second or third degreeburn in a patient, comprising: administering an oxidative reductivepotential water solution to the patient in an amount sufficient to treatthe burn, wherein the oxidative reductive potential water solutioncomprises free chlorine species at a level of from 50 ppm to 80 ppm,wherein the free chlorine species in the solution comprises hypochlorousacid in an amount from 15 ppm to 35 ppm and sodium hypochlorite in anamount from 25 ppm to 50 ppm, wherein the solution is stable for atleast two months, and wherein the pH of the solution is from 7.4 to 7.6.12. The method of claim 11, wherein the solution is administered to thepatient by spraying the burn with the solution.
 13. The method of claim12, wherein the solution is administered to the patient by spraying theburn with the solution with a high pressure irrigation device.
 14. Themethod of claim 12, wherein the burn is moistened with the solution forat least five minutes.
 15. The method of claim 14, wherein the burn ismoistened with the solution for at least 15 minutes.
 16. The method ofclaim 11, wherein the solution is administered to the patient at leastdaily.
 17. The method of claim 16, wherein the solution is administeredto the patient three times a day.
 18. A method of treating a second orthird degree burn in a patient, comprising: (1) spraying the burn withan oxidative reductive potential water solution at high pressure; (2)optionally soaking the burn with an oxidative reductive potential watersolution; (3) spraying the burn with an oxidative reductive potentialwater solution; (4) allowing the oxidative reductive potential watersolution to moisten the burn, wherein the oxidative reductive potentialwater solution has a pH of 7.4 to 7.6 and is stable for at least twomonths, wherein the oxidative reductive potential water solutioncomprises free chlorine species at a level of from 50 ppm to 80 ppm,wherein the free chlorine species is selected from the group selectedfrom the group consisting of hypochlorous acid, hypochlorite ions,sodium hypochlorite, chlorite ions, chloride ions, dissolved chlorinegas, and mixtures thereof; and wherein the free chlorine speciescomprises at least one of: hypochlorous acid in an amount of 15 ppm to35 ppm and sodium hypochlorite in an amount of 25 ppm to 50 ppm.
 19. Themethod of claim 18, wherein the burn is subject to debridement therapyprior to spraying.
 20. The method of claim 1, wherein antibiotics arenot administered to the patient during treatment of the burn.
 21. Themethod of claim 11, wherein antibiotics are not administered to thepatient during treatment of the burn.
 22. The method of claim 18,wherein antibiotics are not administered to the patient during treatmentof the burn.
 23. The method of claim 18, further comprising: applyingskin grafts to the patient.
 24. The method of claim 18, wherein steps(3)-(4) are repeated three times a day.
 25. The method of claim 18,wherein steps (1)-(4) are repeated until the burn is sufficientlyhealed.
 26. The method of claim 18, further comprising: administeringantibiotics to the patient.
 27. The method of claim 1, wherein an extentof burn coverage in the patient that is treated is in a range of fromabove 0% to 19%.
 28. The method of claim 11, wherein an extent of burncoverage in the patient that is treated is in a range of from above 0%to 19%.
 29. The method of claim 18, wherein an extent of burn coveragein the patient that is treated is in a range of from above 0% to 19%.30. The method of claim 5, wherein an extent of burn coverage in thepatient that is treated is in a range of from 30% to 49%.
 31. The methodof claim 11, wherein an extent of burn coverage in the patient that istreated is in a range of from 30% to 49%.
 32. The method of claim 18,wherein an extent of burn coverage in the patient that is treated is ina range of from 30% to 49%.